New RSV RNA molecules and formulations for vaccination

Abstract

A pharmaceutical composition comprising at least one artificial mRNA, wherein the at least one artificial mRNA comprises: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97% identical to SEQ ID NO: 4957, wherein the at least one antigenic peptide or protein is an F protein with a deleted C-terminus (F-del), and wherein the RSV F protein comprises a DSCav1 mutation (S155C, S290C, S190F and V207L), wherein the RSV F protein comprises the two subunits F2 and F1 in a single polypeptide chain, where F2 and F1 are connected via a linker element, where the F2 linker F1 protein aa104-aa144 is missing, wherein the RSV F protein includes at least one further mutation selected from A149C and Y458C, where the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO: 68.

Description

Introduction

[0001] The present invention relates to artificial RNA suitable for use in the treatment or prophylaxis of respiratory syncytial virus (RSV) infection or a disease associated with such infection. In particular, the artificial RNA of the invention comprises at least one heterologous untranslated region (UTR), preferably a 3'-UTR and / or a 5'-UTR, and a coding region encoding at least one antigenic peptide or protein derived from RSV, in particular at least one antigenic peptide or protein derived from the RSV fusion protein (F). The artificial RNA is preferably characterized by increased expression efficiencies of the coding regions operatively linked to the UTR elements.The present invention also relates to compositions and vaccines comprising the artificial RNA in combination with a polymeric carrier, a polycationic protein or peptide, or a lipid nanoparticle (LNP). Furthermore, the invention relates to a kit, in particular a kit of parts, containing the artificial RNA, the composition, or the vaccine. The invention further relates to first and second medical uses of the artificial RNA, the composition, or the vaccine.

[0002] Respiratory syncytial virus (RSV) is an enveloped, non-segmented, negative-sense RNA virus belonging to the family Paramyxoviridae, genus Pneumovirus. It is the most common cause of bronchiolitis and pneumonia in infants under one year of age. RSV also causes recurrent infections, including severe lower respiratory tract infections, which can occur at any age, particularly in the elderly or those with compromised heart, lung, or immune systems. Passive immunization is currently used to prevent severe RSV infections, especially in premature infants, those with bronchopulmonary dysplasia, or congenital heart defects.

[0003] The recommended treatment for RSV bronchiolitis primarily consists of respiratory support and fluid replacement. Specific antiviral therapy is not recommended. The neutralizing monoclonal antibody palivizumab is used for prophylaxis in infants at highest risk of severe infection, but it is too expensive and impractical for general use. Currently, there is no licensed / approved RSV vaccine, and the development of a safe and effective RSV vaccine is a global public health priority.

[0004] In a vaccine trial in the 1960s, infants and young children were immunized with either a formalin-inactivated whole-virus RSV preparation (FIRSV) or a corresponding paramyxovirus preparation (FIPIV). Five percent of the subjects immunized with FI-PIV who subsequently contracted RSV naturally during the next RSV season were hospitalized; 80 percent of those immunized with FIRSV who then contracted RSV were hospitalized, and two children died. This potentiation of RSV infection by vaccination poses a particular challenge for the development of vaccines against RSV infections.

[0005] Therefore, respiratory syncytial virus (RSV) infections represent the largest unmet need for infant vaccines in developed countries, and there is a significant unmet need for infant vaccines worldwide. More than 40 years of effort have not yet resulted in an approved RSV vaccine for humans.

[0006] In addition to the aforementioned humanized monoclonal antibody palivizumab, attenuated live attenuated vaccine viruses have been developed that elicit a strong immune response but are not recommended for use in specific target groups (infants, children, the elderly, and immunocompromised patients). Furthermore, DNA vectors expressing the RSV-F protein, which carries B-cell epitopes, have been used to induce the production of neutralizing antibodies. In this context, WO2008 / 077527 and WO96 / 040945 disclose vectors with DNA sequences encoding the RSV-F protein for use as vaccines. However, the use of DNA as a vaccine can be dangerous due to the unwanted introduction into the genome and may potentially lead to disruption of functional genes and cancer or the formation of anti-DNA antibodies.

[0007] WO2015 / 024668 discloses RNA sequences encoding RSV antigenic peptides and proteins selected from the fusion protein F, glycoprotein G, short hydrophobic protein SH, matrix protein M, nucleoprotein N, large polymerase L, M2-1 protein, M2-2 protein, phosphoprotein P, non-structural protein NS1 or non-structural protein NS2, and an antigenic composition containing protamine-complexed RNA suitable for intradermal administration.

[0008] WO2017 / 070622 discloses a vaccine comprising RNA encoding antigenic RSV peptides and proteins selected from glycoprotein F and glycoprotein G, wherein the RNA is formulated in lipid nanoparticles.

[0009] Apart from some of the approaches mentioned above, there remains an unmet medical need for an effective vaccine for the prophylaxis or treatment of RSV infections.

[0010] Accordingly, the objective of the underlying invention is to provide novel artificial RNA encoding antigenic peptides or proteins of RSV and compositions / vaccines containing this RNA for use as a vaccine for the prophylaxis or treatment of RSV infections, particularly in infants, newborns, pregnant women, elderly and immunocompromised patients.

[0011] Furthermore, it would be desirable for an RNA-based composition or an RNA-based vaccine to have some of the following advantageous features: - Improved translation of RNA constructs at the injection site (e.g., in muscle) - Highly efficient induction of RSV antigen-specific immune responses against the encoded antigenic peptide or protein at very low doses and dosing regimens. - Suitability for immunizing mothers - Suitability for vaccinating infants and / or newborns - Suitable for intramuscular administration - Induction of an RSV-specific functional humoral immune response - Induction of an RSV-specific B-cell memory - Faster onset of immunity against RSV - Longevity of induced immune responses against RSV - Induction of a broad cellular T-cell response against RSV - Induction of a (local and temporary) pro-inflammatory environment - No induction of a systemic cytokine or chemokine response after administration of the vaccine. - Well tolerated, no side effects, non-toxic, - No increase in RSV infection due to vaccination - Advantageous stability properties of the vaccine - Speed, adaptability, simplicity and scalability of RSV vaccine production

[0012] The problems described above are solved by the claimed subject matter. In particular, by a pharmaceutical composition comprising at least one artificial mRNA comprising at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97% identical to SEQ ID NO: 4957, wherein the at least one antigenic peptide or protein is an F protein with a deleted C-terminus (F-del), and wherein the RSV F protein comprises a DSCav1 mutation (S155C, S290C, S190F and V207L), wherein the RSV F protein comprises the two subunits F2 and F1 in a single polypeptide chain,wherein F2 and F1 are linked via a linker element, wherein the F2-linker-F1 protein lacks aa104-aa144, wherein the RSV F protein includes at least one further mutation selected from A149C and Y458C, wherein the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO: 68.

[0013] Furthermore, a pharmaceutical composition comprising at least one artificial mRNA is provided, wherein the at least one artificial mRNA is: (a) a Cap1 structure; (b) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); (c) at least one coding sequence encoding at least one antigenic peptide or protein derived from an RSV fusion (F) protein, wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97%, 98% or 99% identical to SEQ ID NO: 4957, wherein the at least one coding sequence includes at least one nucleic acid sequence that is at least 90%, 91%, 92%, 93% identical to SEQ ID NO: 4958 or 4962; (d) a poly(A) sequence comprising about 100 adenosine nucleotides; wherein all uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ) nucleotides; wherein the at least one artificial mRNA is contained in LNPs.

[0014] Furthermore, a vaccine is provided that includes the pharmaceutical composition, optionally eliciting an adaptive immune response and / or a T-cell immune response against RSV.

[0015] The invention further comprises a kit or kit of parts comprising the pharmaceutical composition and / or the vaccine, optionally comprising a liquid solubilizing agent, and further optionally technical instructions with information on the administration and dosage of the component(s), wherein the components may be the pharmaceutical composition and / or the vaccine.

[0016] Also provided is the pharmaceutical composition, the vaccine, or the components of the kit or kit parts for use as a medicinal product. The pharmaceutical composition, the vaccine, or the components of the kit or kit parts are optionally available for use in the treatment or prophylaxis of RSV infection in a subject or a disorder associated with such infection. Alternatively, the pharmaceutical composition, the vaccine, or the components of the kit or kit parts are available for use in a procedure to reduce the RSV titer in nasal tissue of a subject with RSV infection. Also optionally, the pharmaceutical composition, the vaccine, or the components of the kit or kit parts are available for use in a procedure to reduce the RSV titer in lung tissue of a subject with RSV infection.Preferably, the subject is a human subject, preferably a newborn, a pregnant woman, a breastfeeding woman, an elderly and / or an immunocompromised human subject. Definitions

[0017] For the sake of clarity and readability, the following definitions are given. Every technical feature mentioned in these definitions can be inferred from any single embodiment of the invention. Additional definitions and explanations may be given specifically in connection with these embodiments.

[0018] Percentages in conjunction with numbers are to be understood relative to the total number of the respective items. In other cases, and unless the context specifies otherwise, percentages are to be understood as weight percentages (wt%).

[0019] Adaptive immune response: The term "adaptive immune response," as used here, is recognized and understood by those skilled in the art and refers, for example, to an antigen-specific response of the immune system (the adaptive immune system). Antigen specificity enables the generation of responses tailored to specific pathogens or pathogen-infected cells. The ability to trigger these tailored responses is normally maintained in the body by "memory cells" (B cells). In the invention, the antigen is provided by the coding sequence of artificial RNA that codes for at least one antigenic peptide or protein.

[0020] Antigen: The term “antigen,” as used herein, is recognized and understood by those skilled in the art and refers, for example, to a substance that can be recognized by the immune system, preferably the adaptive immune system, and is capable of triggering an antigen-specific immune response, e.g., by the formation of antibodies and / or antigen-specific T cells as part of an adaptive immune response. An antigen can typically be or comprise a peptide or protein that can be presented by the MHC from T cells. Fragments, variants, and derivatives of peptides or proteins, e.g., those derived from the RSV-F protein and comprising at least one epitope, are also understood as antigens within the scope of the invention. Within the scope of the present invention, an antigen can be the translation product of a provided artificial RNA, as described herein.

[0021] Antigenic peptide or protein: The term “antigenic peptide or protein” is recognized and understood by those skilled in the art and is intended to refer, for example, to a peptide, protein (or polyprotein) derived from an (antigenic) protein / polyprotein that can stimulate the body’s adaptive immune system to mount an adaptive immune response. Therefore, an “antigenic peptide or protein” comprises at least one epitope (as defined herein) or antigen (as defined herein) of the protein from which it is derived (e.g., within the scope of the invention, RSV peptide or protein, preferably RSV-F protein or variants thereof).

[0022] Artificial nucleic acid: The term "artificial nucleic acid," as used here, is recognized and understood by those skilled in the art and refers, for example, to an artificial nucleic acid that does not occur naturally. An artificial nucleic acid can be a DNA molecule, an RNA molecule, or a hybrid molecule with DNA and RNA components. Typically, artificial nucleic acids can be designed and / or produced by genetic engineering techniques to correspond to a desired artificial nucleotide sequence (heterologous sequence). In this context, an artificial sequence is usually a sequence that does not occur in nature, i.e., it differs from the wild-type sequence by at least one nucleotide. The term "wild type" ("wt"), as used here, is recognized and understood by those skilled in the art and refers, for example, to a sequence that occurs in nature.Furthermore, the term “artificial nucleic acid” is not limited to “a single molecule”, but usually encompasses an ensemble of essentially identical molecules.

[0023] Artificial RNA: The term "artificial RNA" as used here refers to RNA that does not occur naturally. In other words, artificial RNA can be understood as a non-natural nucleic acid molecule. Such RNA molecules may be non-natural due to their individual sequence (which does not occur in nature, e.g., G / C ratio, modified coding sequence, UTRs) and / or due to other modifications, e.g., structural modifications of nucleotides that do not occur in nature. Typically, artificial RNA can be designed and / or produced by genetic engineering to match a desired artificial nucleotide sequence (heterologous sequence). In this context, an artificial RNA sequence is generally a sequence that does not occur in nature; that is, it differs from the wild-type sequence by at least one nucleotide.The term “artificial RNA” is not limited to “a single molecule” but generally encompasses an ensemble of substantially identical molecules. Accordingly, it can refer to a multitude of substantially identical RNA molecules contained in an aliquot or sample. In the context of the invention, the RNA of the invention is artificial RNA as defined herein.

[0024] Cationic: Unless otherwise specified by the context, the term "cationic" means that the structure in question carries a positive charge, either permanently or not permanently, depending on certain conditions such as pH. Thus, the term "cationic" encompasses both "permanently cationic" and "cationizable".

[0025] Cationizable: The term "cationizable" as used here means that a compound, group, or atom is positively charged at a lower pH and uncharged at a higher pH of the environment. Even in non-aqueous environments where pH cannot be determined, a cationizable compound, group, or atom is positively charged at a high hydrogen ion concentration and uncharged at a low concentration or activity of hydrogen ions. The pH or hydrogen ion concentration at which it is charged or uncharged depends on the individual properties of the cationizable or polycationizable compound, in particular the pKa of the respective cationizable group or atom.In a dilute aqueous environment, the proportion of cationizable compounds, groups, or atoms carrying a positive charge can be estimated using the so-called Henderson-Hasselbalch equation, which is known to those skilled in the art. In some embodiments, for example, it is preferred that if a compound or unit is cationizable, it is positively charged at a pH of about 1 to 9, preferably 4 to 9, 5 to 8, or even 6 to 8, more preferably at a pH of or below 9, 8, or 7, and most preferably at physiological pH values, e.g., about 7.3 to 7.4, i.e., under physiological conditions, particularly under physiological salt conditions of the cell in vivo. In other embodiments, it is preferred that the cationizable compound or fraction is positively charged at physiological pH values, e.g.,The pH is approximately 7.0 to 7.4, predominantly neutral, but becomes positively charged at lower pH values. In some embodiments, the preferred pKa range for the cationizable compound or fraction is approximately 5 to approximately 7.

[0026] Coding sequence / coding region: The terms "coding sequence" or "coding region" and the corresponding abbreviation "cds", as used herein, are recognized and understood by those skilled in the art and are intended to refer, for example, to a sequence of several nucleotide triplets that can be translated into a peptide or protein. A coding sequence within the meaning of the present invention is preferably an RNA sequence consisting of a number of nucleotides divisible by three, beginning with a start codon and preferably ending with a stop codon.

[0027] Composition: In the context of the invention, a “composition” refers to any type of composition into which the specified ingredients (e.g., artificial RNA of the invention in combination with LNP) can be incorporated, optionally together with other components, generally with at least one pharmaceutically acceptable carrier or excipient. The composition may therefore be a dry composition such as a powder or granules, or a solid unit such as a lyophilized form or a tablet. Alternatively, the composition may be in liquid form, with each component being incorporated independently in dissolved or dispersed (e.g., suspended or emulsified) form.

[0028] Compound: As used herein, a “compound” is a chemical substance, which is a material composed of molecules that have essentially the same chemical structure and properties. In a small-molecule compound, the molecules are typically identical with respect to their atomic composition and structural configuration. In a macromolecular or polymeric compound, the molecules of a compound are very similar, but not all are necessarily identical. For example, a segment of a polymer that is supposed to consist of 50 monomer units may also contain individual molecules with, for example, 48 or 53 monomer units.

[0029] Derived from: The term “derived from”, as used in the present description in connection with a nucleic acid, i.e. for a nucleic acid “derived from” another nucleic acid, means that the nucleic acid derived from another nucleic acid has at least 50%, preferably at least 55%, preferably at least 60%, preferably at least 65%, preferably at least 70%, more preferably at least 75%, more preferably at least 80%, 81%, 82%, 83%, 84%, more preferably at least 85%, 86%, 87%, 88%, 89%, more preferably at least 90%, 91%, 92%, 93%, 94%, more preferably at least 95%, 96%, 97% and particularly preferably at least 98%, 99% sequence identity with the nucleic acid from which it is derived. It is known to those skilled in the art that sequence identity is typically calculated for the same types of nucleic acids, i.e., for DNA sequences or for RNA sequences.Therefore, if DNA is "derived" from RNA, or if RNA is "derived" from DNA, the first step involves converting the RNA sequence into the corresponding DNA sequence (in particular by replacing the uracil (U) residues with thymidine (T) residues throughout the sequence), or vice versa, converting the DNA sequence into the corresponding RNA sequence (in particular by replacing the thymidine (T) residues with uracil (U) residues throughout the sequence). Subsequently, the sequence identity of the DNA sequences or the sequence identity of the RNA sequences is determined. Preferably, a nucleic acid "derived" from a nucleic acid is also a nucleic acid that is modified compared to the nucleic acid from which it is derived, for example, to further increase RNA stability and / or to extend and / or increase protein production.It goes without saying that modifications are preferred which do not impair the stability of the RNA, e.g. compared to the nucleic acid from which it is derived. In the context of amino acid sequences (e.g., antigenic peptides or proteins), the term "derived from" means that the amino acid sequence derived from another amino acid sequence (e.g., RSV F protein) has at least 50%, preferably at least 55%, preferably at least 60%, preferably at least 65%, preferably at least 70%, more preferably at least 75%, more preferably at least 80%, 81%, 82%, 83%, 84%, more preferably at least 85%, 86%, 87%, 88%, 89%, even more preferably at least 90%, 91%, 92%, 93%, 94%, even more preferably at least 95%, 96%, 97% and particularly preferably at least 98%, 99% sequence identity with the amino acid sequence from which it is derived.

[0030] Therefore, if an antigenic peptide or protein is “derived” from an RSV fusion protein (F), it is understood that the antigenic peptide or protein “derived” from the RSV-F protein is a variant or fragment of the RSV-F protein, e.g., F0 (full-length precursor), F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F-del,DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3 ​​(as indicated here). Furthermore, the antigenic peptides or proteins "derived" from RSV-F proteins (e.g., F0, F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F-del_DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3) may differ in their amino acid sequence, exhibiting a certain percentage of identity as defined above. Suitable additional examples of RSV-F proteins from which an antigenic peptide or protein may be "derived" are listed in Table 1.

[0031] Epitope: The term "epitope" (also called "antigenic determinant" in technical jargon), as used here, is recognized and understood by those skilled in the art and refers, for example, to T-cell epitopes and B-cell epitopes. T-cell epitopes or parts of the antigenic peptides or proteins can comprise fragments with a length of preferably about 6 to about 20 or even more amino acids, e.g., fragments such as those processed and presented by MHC class I molecules, preferably with a length of about 8 to about 10 amino acids, e.g., 8, 9, or 10 (or even 11 or 12 amino acids), or fragments such as those processed and presented by MHC class II molecules, preferably with a length of about 13 to about 20 or even more amino acids, wherein these fragments can be selected from any part of the amino acid sequence.These fragments are typically recognized by T cells as a complex consisting of the peptide fragment and an MHC molecule; that is, the fragments are not normally recognized in their native form. B-cell epitopes are typically fragments located on the outer surface of (native) protein or peptide antigens, preferably with 5 to 15 amino acids, more preferably with 5 to 12 amino acids, and still more preferably with 6 to 9 amino acids, which can be recognized by antibodies, i.e., in their native form. Furthermore, such protein or peptide epitopes can be selected from any of the variants of such proteins or peptides mentioned herein.In this context, antigenic determinants can be conformational or discontinuous epitopes consisting of segments of the proteins or peptides defined herein, which are discontinuous in their amino acid sequence but are joined together in their three-dimensional structure, or continuous or linear epitopes consisting of a single polypeptide chain. In the context of the present invention, an epitope can be the translation product of a provided artificial RNA, as described herein.

[0032] Fragment: The term "fragment," as used in this description in connection with a nucleic acid sequence or an amino acid sequence, can typically be a shorter segment of a full-length sequence, such as a nucleic acid sequence or an amino acid sequence. Accordingly, a fragment typically consists of a sequence that is identical to the corresponding segment within the full-length sequence.A preferred fragment of a sequence within the scope of the present invention consists of a continuous section of units, such as nucleotides or amino acids, corresponding to a continuous section of units in the molecule from which the fragment is derived, representing at least 5%, 10%, 20%, preferably at least 30%, more preferably at least 40%, more preferably at least 50%, more preferably at least 60%, more preferably at least 70% and most preferably at least 80% of the total (i.e. full length) molecule from which the fragment is derived (e.g. RSV F-protein).The term "fragment," as used in this description in connection with proteins or peptides, typically comprises a sequence of a protein or peptide, as defined herein, which is N-terminally and / or C-terminally truncated with respect to its amino acid sequence (or its encoded nucleic acid molecule) compared to the amino acid sequence of the original (native) protein (or its encoded nucleic acid molecule). Such truncation may therefore occur either at the amino acid level or, correspondingly, at the nucleic acid level. Sequence identity with respect to such a fragment, as defined herein, may therefore preferably refer to the entire protein or peptide, as defined herein, or to the entire (encoding) nucleic acid molecule of such a protein or peptide. In the context of antigens, such a fragment may have a length of approximately 6 to approximately 20 or even more amino acids, e.g.,Fragments as processed and presented by MHC class I molecules, preferably with a length of about 8 to about 10 amino acids, e.g., 8, 9, or 10 (or even 6, 7, 11, or 12 amino acids), or fragments as processed and presented by MHC class II molecules, preferably with a length of about 13 or more amino acids, e.g., 13, 14, 15, 16, 17, 18, 19, 20, or even more amino acids, wherein these fragments can be selected from any part of the amino acid sequence. These fragments are typically recognized by T cells in the form of a complex consisting of the peptide fragment and an MHC molecule; that is, the fragments are typically not recognized in their native form. Fragments of proteins or peptides (e.g., in connection with antigens) can contain at least one epitope of these proteins or peptides.Furthermore, domains of a protein, such as the extracellular domain, the intracellular domain or the transmembrane domain, as well as shortened or truncated versions of a protein, can also be understood as fragments of a protein.

[0033] Heterologous: The terms "heterologous" or "heterologous sequence," as used in this description in connection with a nucleic acid sequence or an amino acid sequence, refer to a sequence (e.g., DNA, RNA, amino acid) as recognized and understood by a person skilled in the art, and are intended to refer to a sequence derived from a different gene, allele, or species. Two sequences are generally described as "heterologous" if they cannot be derived from the same gene or allele. That is, heterologous sequences may originate from the same organism, but do not naturally occur (in nature) in the same nucleic acid molecule, such as the same RNA or protein.

[0034] Humoral immune response: The terms "humoral immunity" or "humoral immune response" are recognized and understood by experts and refer, for example, to B-cell-mediated antibody production and, where applicable, to additional processes that accompany antibody production. A humoral immune response can typically be characterized, for example, by Th2 activation and cytokine production, germinal center formation and isotype switching, affinity maturation, and memory cell formation. Humoral immunity can also typically refer to the effector functions of antibodies, which include the neutralization of pathogens and toxins, classical complement activation, and the promotion of phagocytosis and pathogen elimination by opsonins.

[0035] Identity (of a sequence): The term "identity," as used in this description in connection with a nucleic acid sequence or an amino acid sequence, is recognized and understood by those skilled in the art and refers, for example, to the percentage to which two sequences are identical. To determine the percentage to which two sequences are identical, e.g., nucleic acid sequences or amino acid sequences as defined herein, preferably the amino acid sequences encoded by the artificial nucleic acid sequence as defined herein, or the amino acid sequences themselves, the sequences can be aligned and subsequently compared. For example, a position of a first sequence can be compared with the corresponding position of the second sequence.If a position in the first sequence contains the same component (remainder) as a position in the second sequence, the two sequences are identical at that position. If this is not the case, the sequences differ at that position. If insertions occur in the second sequence compared to the first, gaps can be inserted into the first sequence to allow for further alignment. If deletions occur in the second sequence compared to the first sequence, gaps can be inserted into the second sequence to allow for further alignment. The percentage to which two sequences are identical is then a function of the number of identical positions divided by the total number of positions, including positions that are occupied in only one sequence. The percentage to which two sequences are identical can be determined using a mathematical algorithm.A preferred, but not limited, example of a mathematical algorithm that may be used is the algorithm integrated into the BLAST program. This program can identify sequences that are identical to some extent to the sequences of the present invention.

[0036] Immunogen, immunogen: The terms "immunogen" or "immunogenic" are recognized and understood by those skilled in the art and are intended to refer, for example, to a compound that can stimulate / induce an immune response. Preferably, an immunogen is a peptide, polypeptide, or protein. An immunogen within the meaning of the present invention is the translation product of a provided artificial nucleic acid, preferably RNA, comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, as defined herein. Typically, an immunogen elicits an adaptive immune response.

[0037] Immune response: The term "immune response" is recognized and understood by experts and refers, for example, to a specific response of the adaptive immune system to a particular antigen (so-called specific or adaptive immune response) or to a non-specific response of the innate immune system (so-called non-specific or innate immune response) or to a combination thereof.

[0038] Immune system: The term "immune system" is recognized and understood by experts and refers, for example, to a system within an organism that can protect it from infections. If a pathogen manages to overcome an organism's physical barrier and enter it, the innate immune system mounts an immediate but non-specific response. If pathogens evade this innate response, vertebrates possess a second line of defense: the adaptive immune system. Here, the immune system adjusts its response during an infection to improve pathogen recognition. This enhanced response is then retained after the pathogen is eliminated in the form of immunological memory, enabling the adaptive immune system to mount faster and stronger attacks upon any subsequent encounter with the pathogen.According to this view, the immune system consists of the innate and the adaptive immune system. Each of these two parts typically contains so-called humoral and cellular components.

[0039] Innate immune system: The term "innate immune system" (also known as the non-specific or innate immune system) is recognized and understood by experts and refers, for example, to a system that typically includes the cells and mechanisms that protect the host in a non-specific way from infections by other organisms. This means that the cells of the innate system can recognize pathogens and generally respond to them, but unlike the adaptive immune system, it does not confer lasting or protective immunity on the host. The innate immune system can, for example, B. by ligands of Toll-like receptors (TLRs) or other excipients such as lipopolysaccharides, TNF-alpha, CD40 ligand or cytokines, monokines, lymphokines, interleukins or chemokines, IL-1 to IL-33, IFN-alpha, IFN-beta, IFN-gamma, GM-CSF, G-CSF, M-CSF, LT-beta, TNF-alpha, growth factors and hGH, a ligand of the human Toll-like receptor (zg, TLR1 to TLR10), a ligand of a murine Toll-like receptor (e.g. TLR1 to TLR13), a ligand of a NOD-like receptor, a ligand of a RIG-I-like receptor, an immunostimulatory nucleic acid, an immunostimulatory RNA (isRNA), a CpG-DNA, an antibacterial agent or an antiviral agent may be activated.

[0040] Lipidoid compound: A lipidoid compound, also simply called a lipidoid, is a lipid-like compound, i.e., an amphiphilic compound with lipid-like physical properties. Within the scope of the present invention, the term lipid also includes lipidoid compounds.

[0041] Monovalent vaccine, monovalent composition: The terms “monovalent vaccine,” “monovalent composition,” “univalent vaccine,” or “univalent composition” are recognized and understood by those skilled in the art and are intended to refer, for example, to a composition or vaccine containing only one antigen of a virus. Accordingly, the vaccine or composition comprises only one RNA species encoding a single antigen for a single organism. The term “monovalent vaccine” encompasses immunization against a single valence. Within the scope of the invention, a monovalent RSV vaccine or composition would comprise an artificial RNA encoding a single antigenic peptide or protein derived from a specific RSV (e.g., RSV F).

[0042] Nucleic acid: The terms "nucleic acid" or "nucleic acid molecule" are recognized and understood by those skilled in the art and are intended to refer, for example, to a molecule that comprises, or preferably consists of, nucleic acid components. The term "nucleic acid molecule" preferably refers to DNA or RNA molecules. It is preferably used synonymously with the term polynucleotide. Preferably, a nucleic acid or nucleic acid molecule is a polymer comprising or consisting of nucleotide monomers covalently linked by phosphodiester bonds of a sugar / phosphate backbone. The term "nucleic acid molecule" also includes modified nucleic acid molecules, such as base-, sugar-, or scaffold-modified DNA or RNA molecules, as defined herein.

[0043] Nucleic acid sequence / RNA sequence / amino acid sequence: The terms "nucleic acid sequence", "RNA sequence" or "amino acid sequence" are recognized and understood by experts and are intended to refer, for example, to a specific and individual sequence of nucleotides or amino acids.

[0044] Permanently cationic: The term "permanently cationic," as used here, is recognized and understood by those skilled in the art and means, for example, that the compound, group, or atom in question is positively charged at every pH value or hydrogen ion activity of its environment. Typically, the positive charge results from the presence of a quaternary nitrogen atom. If a compound carries a multitude of such positive charges, it can be described as permanently polycationic, which is a subcategory of permanently cationic.

[0045] Pharmaceutically effective amount: The terms “pharmaceutically effective amount” or “effective amount” are recognized and understood by those skilled in the art and are intended to refer, for example, to an amount of a compound (e.g., the artificial RNA of the invention) sufficient to produce a pharmaceutical effect, such as, in connection with the invention, an immune response (e.g., against an antigenic peptide, protein, polyprotein, as defined herein).

[0046] Polyvalent / multivalent vaccine, polyvalent / multivalent composition: The terms "polyvalent vaccine," "polyvalent composition," "multivalent vaccine," or "multivalent composition" are recognized and understood by those skilled in the art and are intended to refer, for example, to a composition or vaccine containing antigens from more than one virus strain, or different antigens of the same virus, or any combination thereof. These terms describe the fact that the vaccine or composition has more than one valence. Within the scope of the invention, a polyvalent RSV vaccine would comprise a vaccine comprising artificial RNA encoding antigenic peptides or proteins derived from several different RSV strains, or artificial RNA encoding different antigens of the same RSV strain, or a combination thereof.In a preferred embodiment, a polyvalent RSV vaccine or a polyvalent RSV composition comprises more than one, preferably two, three, four, or even more, different artificial RNA species, each encoding at least one different antigenic peptide or protein of RSV (e.g., RSV F and RSV M or RSV F and RSV N). Methods for producing polyvalent mRNA vaccines are disclosed in PCT application PCT / EP2016 / 082487 or in published patent application WO2017 / 1090134A1.

[0047] Stabilized nucleic acid molecule or stabilized RNA: The term "stabilized nucleic acid molecule" or "stabilized RNA" refers to a nucleic acid molecule, preferably an RNA molecule, that is modified to be more stable against degradation or breakdown, e.g., by environmental factors or enzymatic digestion, such as exo- or endonuclease degradation, than the nucleic acid molecule without the modification. Preferably, within the scope of the present invention, a stabilized nucleic acid molecule, e.g., stabilized RNA, is stabilized in a cell, such as a prokaryotic or eukaryotic cell, preferably in a mammalian cell, such as a human cell. The stabilization effect can also be exerted outside of cells, e.g., in a buffer solution, etc., e.g., in a manufacturing process for a pharmaceutical composition containing the stabilized nucleic acid molecule.

[0048] T-cell responses: The terms "cellular immunity," "cellular immune response," or "cellular T-cell responses" used here are recognized and understood by experts and refer, for example, to the activation of macrophages, natural killer (NK) cells, antigen-specific cytotoxic T lymphocytes, and the release of various cytokines in response to an antigen. More generally, cellular immunity is not based on antibodies but on the activation of immune system cells. Typically, a cellular immune response can be characterized, for example, by the activation of antigen-specific cytotoxic T lymphocytes, which are capable of inducing apoptosis in cells, such as specific immune cells like dendritic cells or other cells that display epitopes of foreign antigens on their surface.Within the scope of the invention, the antigen is provided by artificial RNA encoding at least one antigenic peptide or protein derived from RSV, which suitably induces T-cell responses. The artificial RNA, composition, and vaccine of the invention advantageously elicit cellular T-cell responses against RSV F antigens.

[0049] Variant (of a sequence): The term "variant," as used in this description in connection with a nucleic acid sequence, is recognized and understood by those skilled in the art and refers, for example, to a variant of nucleic acid sequences that forms the basis of a nucleic acid sequence. A variant nucleic acid sequence may, for example, have one or more nucleotide deletions, insertions, additions, and / or substitutions compared to the nucleic acid sequence from which the variant is derived. Preferably, a variant of a nucleic acid sequence is identical to the nucleic acid sequence from which the variant is derived to at least 40%, preferably at least 50%, more preferably at least 60%, more preferably at least 70%, even more preferably at least 80%, even more preferably at least 90%, and most preferably at least 95%. Preferably, the variant is a functional variant.A “variant” of a nucleic acid sequence can exhibit at least 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% nucleotide identity over a section of 10, 20, 30, 50, 75 or 100 nucleotides of such nucleic acid sequence.

[0050] The term “variant,” as used in the present description in connection with proteins or peptides, is recognized and understood by those skilled in the art and is intended to refer, for example, to a protein or peptide variant with an amino acid sequence that differs from the original sequence by one or more mutations, such as one or more substituted, inserted, and / or deleted amino acids. Preferably, these fragments and / or variants have the same biological function or specific activity as the native full-length protein, e.g., its specific antigenic property. “Variants” of proteins or peptides, as defined within the scope of the present invention, may include conservative amino acid substitutions compared to their native, i.e., unmutated, physiological sequence.These amino acid sequences, as well as their coding nucleotide sequences, fall particularly under the concept of variants as defined here. Substitutions in which amino acids from the same class are exchanged for one another are called conservative substitutions. These are, in particular, amino acids with aliphatic side chains, positively or negatively charged side chains, aromatic groups in the side chains, or amino acids whose side chains can form hydrogen bonds, e.g., side chains with a hydroxyl function. This means, for example, that an amino acid with a polar side chain is replaced by another amino acid with a similarly polar side chain, or, for example, an amino acid characterized by a hydrophobic side chain is replaced by another amino acid with a similarly hydrophobic side chain (e.g., serine (threonine) by threonine (serine) or leucine (isoleucine) by isoleucine (leucine)).Insertions and substitutions are possible, in particular, at sequence positions that do not alter the three-dimensional structure or affect the binding region. Changes in the three-dimensional structure due to insertion(s) or deletion(s) can be easily detected, for example, using circular dichroism (CD) spectra. A "variant" of a protein or peptide can exhibit at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% amino acid identity over a segment of 10, 20, 30, 50, 75, or 100 amino acids of such protein or peptide. Preferably, a variant of a protein is a functional variant of the protein, meaning that the variant exerts the same effect or functionality as the protein from which it is derived.

[0051] 3' untranslated region, 3' UTR element, 3' UTR: The term "3' untranslated region" or "3' UTR element" is recognized and understood by those skilled in the art and is intended to refer, for example, to a part of a nucleic acid molecule that is located 3' (i.e., "downstream") from a coding sequence and is not normally translated into a protein. Normally, a 3' UTR is the part of an mRNA that lies between the coding sequence (cds) and the poly(A) sequence of the mRNA. Within the scope of the invention, the term 3' UTR may also include elements that are not encoded in the DNA template from which an artificial RNA is transcribed, but which are added after transcription during maturation, e.g., a poly(A) sequence.

[0052] 5' untranslated region, 5' UTR element, 5' UTR: The term "5' untranslated region (5' UTR)" is recognized and understood by experts and is said to refer, for example, to a part of a nucleic acid molecule that is located 5' (i.e., "upstream") from a coding sequence and is not translated into protein. A 5' UTR is generally understood to be a specific segment of messenger RNA (mRNA) located 5' from the coding sequence of the mRNA. Typically, the 5' UTR begins at the transcription start site and ends one nucleotide before the start codon of the coding sequence. Preferably, 5' UTRs are longer than 20, 30, 40, or 50 nucleotides. The 5' UTR may contain elements that control gene expression, also called regulatory elements. Such regulatory elements can include, for example, the 5'-nucleotide polymorphism (5'-UTR), the 5'-nucleotide polymorphism (5'-UTR), and the 5'-nucleotide polymorphism (5'-UTR). These could be ribosomal binding sites. The 5' UTR may be post-transcriptionally modified, e.g., by adding a 5' cap.

[0053] 5'-terminal oligopyrimidine tract (TOP), TOP-UTR: The term "5'-terminal oligopyrimidine tract (TOP)" refers to a segment of pyrimidine nucleotides located in the 5'-terminal region of a nucleic acid molecule, such as the 5'-terminal region of certain mRNA molecules or the 5'-terminal region of a functional unit, e.g., the transcribed region of certain genes. The sequence begins with a cytidine, which usually corresponds to the transcription start site, and is followed by a segment of typically about 3 to 30 pyrimidine nucleotides. The TOP can, for example, contain 3 to 30 or even more nucleotides. The pyrimidine segment, and thus the 5'-TOP, terminates one nucleotide 5' upstream of the first purine nucleotide located downstream of the TOP. Messenger RNA (mRNA) containing a 5'-terminal oligopyrimidine tract is often referred to as TOP mRNA.Accordingly, genes that provide such messenger RNAs are called TOP genes. The term "TOP motif" or "5'-TOP motif" refers to a nucleic acid sequence corresponding to a 5'-TOP as defined above. A TOP motif within the meaning of the present invention is therefore preferably a segment of pyrimidine nucleotides with a length of 3-30 nucleotides. Preferably, the TOP motif consists of at least 3 pyrimidine nucleotides, preferably at least 4 pyrimidine nucleotides, preferably at least 5 pyrimidine nucleotides, more preferably at least 6 nucleotides, more preferably at least 7 nucleotides, and most preferably at least 8 pyrimidine nucleotides, wherein the segment of the pyrimidine nucleotides preferably begins at its 5' end with a cytosine nucleotide.In TOP genes and TOP mRNAs, the TOP motif preferably begins at its 5' end with the transcription start site and terminates one nucleotide 5' upstream of the first purine residue in that gene or mRNA. A TOP motif according to the present invention is preferably located at the 5' end of a sequence that represents a 5' UTR or at the 5' end of a sequence that encodes a 5' UTR. Thus, a segment of 3 or more pyrimidine nucleotides is preferably referred to as a "TOP motif" according to the present invention if this segment is located at the 5' end of a corresponding sequence, such as the artificial nucleic acid, the 5' UTR element of the artificial nucleic acid, or the nucleic acid sequence derived from the 5' UTR of a TOP gene as described herein.In other words, a segment of 3 or more pyrimidine nucleotides that is not located at the 5' end of a 5' UTR or 5' UTR element, but rather somewhere within a 5' UTR or 5' UTR element, is preferably not referred to as a "TOP motif." In some embodiments, the nucleic acid sequence of the 5' UTR element derived from a 5' UTR of a TOP gene terminates at its 3' end with a nucleotide located at position 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 upstream of the start codon (e.g., A(U / T)G) of the gene or RNA from which it is derived. Thus, the 5' UTR element does not comprise any part of the protein-coding sequence. Preferably, the only protein-coding part of the at least one nucleic acid sequence, in particular the RNA sequence, is provided by the coding sequence. Brief description of the invention

[0054] The present invention is based on the inventor's surprising discovery that at least one peptide or protein derived from a respiratory syncytial virus (RSV) F protein encoded by the invention's artificial RNA can be efficiently expressed in a mammalian cell. Even more unexpectedly, the inventors demonstrated that the invention's artificial RNA can induce specific functional and protective immune responses in, for example, cotton rats (see, for example, Examples 2 and 3). Various optimizations in the design of the RSV F antigen further enhanced these immune responses. Furthermore, the expression of the RSV F antigen encoded by the artificial nuclein RNA could be increased by selecting suitable heterologous S' untranslated regions (UTRs) and suitable heterologous 3' untranslated regions (UTRs) (see, for example, Example 4).Advantageously, the artificial RNA according to the invention, which comprises advantageous 3'-UTR / 5'-UTR combinations, induces highly efficient antigen-specific immune responses against the encoded RSV F. Furthermore, the artificial RNA according to the invention, which is contained in lipid nanoparticles (LNPs), highly efficiently induces antigen-specific immune responses against RSV F at very low doses and dosing regimens (see, for example, Example 3). In addition, for example, Example 8 and Example 12 provide compositions / vaccines containing another artificial RNA encoding a further antigen, wherein the artificial RNA encoding a further antigen appropriately elicits or enhances T-cell responses and leads to a Th1-directed immune response, which is considered an important prerequisite for a potential RSV vaccine (Th2-directed responses have been associated in animal models of enhanced respiratory disease (ERD)).Furthermore, the compositions are suitable for inducing T-cell responses. Accordingly, the artificial RNA and the composition / vaccine comprising this artificial RNA of the invention are suitable for triggering an immune response against RSV F in a mammal. The artificial RNA and the composition / vaccine containing the artificial RNA are therefore suitable for use as a vaccine, e.g., as a human vaccine, for example, as a vaccine for pregnant women or infants.

[0055] In a first aspect, an artificial nucleic acid, preferably an artificial RNA, is provided, comprising at least one 5' untranslated region (UTR) and / or at least one 3' untranslated region (UTR) and at least one coding sequence that is operatively linked to the 3' UTR and / or 5' UTR and encodes at least one antigenic peptide or protein derived from the RSV F protein or a fragment or variant thereof. Specifically, a pharmaceutical composition comprising at least one artificial mRNA is protected, wherein the at least one artificial mRNA includes: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97% identical to SEQ ID NO: 4957, wherein at least one antigenic peptide or protein is an F-protein with a deleted C-terminus (F-del), and the RSV F protein includes a DSCav1 mutation (S155C, S290C, S190F and V207L), wherein the RSV F-protein comprises the two subunits F2 and F1 in a single polypeptide chain, F2 and F1 being linked via a linker element, where the F2 linker F1 protein aa104-aa144 is missing, wherein the RSV F protein includes at least one further mutation selected from A149C and Y458C, wherein the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO: 68.

[0056] The protected artificial nucleic acid or artificial RNA of the invention is an artificial mRNA and furthermore as defined in the claims. When reference is made herein to the "artificial RNA of the invention," this refers to the "artificial mRNA of the claimed pharmaceutical composition."

[0057] The artificial mRNA may include at least one nucleic acid sequence derived from a 3' UTR of a gene selected from an ALB7 gene, an alpha globin gene, a PSMB3, CASP1, COX6B1, GNAS, NDUFA1 and RPS9 gene, or from a homolog, fragment or variant of any of these genes.

[0058] In preferred embodiments, the artificial mRNA comprises at least one nucleic acid sequence derived from a 5' UTR of a gene selected from an RPL32 gene, an HSD17B4, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B and UBQLN2 gene, or from a homolog, fragment or variant of any of these genes.

[0059] Advantageously, the artificial mRNA of the invention comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from an RSV F protein and defined according to the claims, which is operatively linked to a 3'-UTR and a 5'-UTR composed of a-1 (HSD17B4 / PSMB3), a-2 (Ndufa4 / PSMB3), a-3 (SLC7A3 / PSMB3), a-4 (NOSIP / PSMB3), a-5 (MP68 / PSMB3), b-1 (UBQLN2 / RPS9), b-2 (ASAH1 / RPS9), b-3 (HSD17B4 / RPS9), b-4 (HSD17B4 / CASP1), b-5 (NOSIP / COX6B1), c-1 (NDUFA4 / RPS9), c-2 (NOSIP / NDUFA1), c-3 (NDUFA4 / COX6B1), c-4 (NDUFA4 / NDUFA1), c-5 (ATP5A1 / PSMB3), d-1 (Rpl31 / PSMB3), d-2 (ATP5A1 / CASP1), d-3 (SLC7A3 / GNAS), d-4 (HSD17B4 / NDUFA1), d-5 (Slc7a3 / Ndufa1), e-1 (TUBB4B / RPS9), e-2 (RPL31 / RPS9), e-3 (MP68 / RPS9), e-4 (NOSIP / RPS9), e-5 (ATP5A1 / RPS9), e-6 (ATP5A1 / COX6B1), f-1 (ATP5A1 / GNAS), f-2 (ATP5A1 / NDUFA1), f-3 (HSD17B4 / COX6B1), f-4 (HSD17B4 / GNAS), f-5 (MP68 / COX6B1), g-1 (MP68 / NDUFA1), g-2 (NDUFA4 / CASP1), g-3 (NDUFA4 / GNAS), g-4 (NOSIP / CASP1),g-5 (RPL31 / CASP1), h-1 (RPL31 / COX6B1), h-2 (RPL31 / GNAS), h-3 (RPL31 / NDUFA1), h-4 (SIc7a3 / CASP1), h-5 (SLC7A3 / COX6B1), i-1 (SLC7A3 / RPS9), i-2 (RPL32 / ALB7), or i-3 (α-globin gene), wherein a-1 (HSD17B4 / PSMB3), a-4 (NDUFA4 / PSMB3), c-1 (NDUFA4 / RPS9), e-4 (NOSIP / RPS9), g-2 (NDUFA4 / CASP1), i-2 (RPL32 / ALB7) or i-3 (alpha-globin) are particularly preferred.

[0060] The at least one antigenic peptide or protein derived from the RSV-F protein and defined according to the claims is an F-protein with a deleted C-terminus (referred to as “F-del”, aa 1-553).

[0061] The at least one antigenic peptide or protein additionally contains a mutation that stabilizes the antigen in the state of preconformation / prefusion conformation, namely a DSCav1 mutation (S155C, S290C, S190F and V207L) (referred to as “DSCav1”, e.g. “F0_DSCav1” or “F-del_DSCav1”).

[0062] The at least one antigenic peptide or protein is a fusion protein comprising the two subunits F1 and F2 of mature F in a single chain, linked via a linker (GS) to increase the stability of the protein (F(1-103)-GS-F(145-574); F(1-103)-GS-F(145-553)).

[0063] The protein comprising the two subunits of mature F in a single chain (referred to as "F2-linker-F1"), e.g. (F(1-103)-GS-F(145-574); F(1-103)-GS-F(145-553), may additionally contain a DSCav1 mutation (referred to here as "mut0").

[0064] The RSV F protein, which comprises the two subunits of mature F in a single chain (referred to as "F2-linker F1"), contains at least one further mutation selected from A149C and Y458C (referred to here as "mut4") and may, in addition to the DSCav1 mutation, include at least one further mutation promoting disulfide bonds between the protomers, the mutations being selectable from (S46G, A149C, S215P, Y458C, K465Q; referred to here as "mut1"), (S46G, E92D, A149C, S215P, Y458C, K465Q; referred to here as "mut2"), or (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q; referred to here as "mut3"), (N183GC, N428C; referred to here as “mut5”), (Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q; referred to here as “mut6”), (Q98C, Q361C, L95M, I221M, R429K; referred to here as “mut7”) or (N183GC, N428C, S46G, N67I, E92D, S21P, K465Q; referred to here as “mut8”).

[0065] The at least one antigenic peptide or protein derived from the RSV-F protein encoded by the artificial RNA of the invention is selected from F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, as described in the claims.

[0066] The at least one coding sequence can encode at least one of the amino acid sequences that are identical or at least 97%, 98% or 99% identical to one of the SEQ ID NO: : 2005, 2743, 3481, 4957, 16523.

[0067] Preferably, the artificial RNA may comprise a coding sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to one of the SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, 21402.

[0068] The artificial RNA may comprise a codon-modified coding sequence selected from a C-maximized coding sequence, a CAI-maximized coding sequence, a coding sequence adapted to human codon use, a G / C content-modified coding sequence, and a G / C-optimized coding sequence, or any combination thereof.

[0069] The artificial RNA can be mRNA, viral RNA, self-replicating RNA, circular RNA, or replicon RNA. In preferred embodiments, the artificial RNA is mRNA.

[0070] The artificial RNA, preferably mRNA, may furthermore comprise at least one selected from a cap structure, a poly(A) sequence, a poly(C) sequence, a histone stem loop and / or a 3'-terminal sequence element.

[0071] The artificial RNA as defined in the claims comprises or preferably consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2014-2373, 21436-21438, 21582-21584, or a fragment or variant thereof (encoding F-del_DSCav1_mut1), SEQ ID NOs: 2752-3111, 21442-21444, 21588-21590 or a fragment or variant thereof (encoding F-del_DSCav1_mut1) F-del_DSCav1_mut2 encoding), SEQ ID NOs: 3490-3849, 21448-21450, 21594-21596, or a fragment or variant thereof (for F-del_DSCav1_mut3 ​​encoding), SEQ ID NOs: 4966-5325, 16532-16891, 21454-21456, 21600-21602, 21526-21528, 21672-21674 or a fragment or variant thereof (for F-del_DSCav1_mut4 encoding).

[0072] In a second aspect, the present invention provides a composition containing the artificial mRNA of the first aspect, as defined in the claims.

[0073] In preferred embodiments, the composition containing the artificial mRNA as defined in the claims comprises at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, selected from matrix protein M, nucleoprotein N, M2-1 protein and / or phosphoprotein P or combinations thereof.

[0074] The matrix protein M, the nucleoprotein N, the M2-1 protein, the M2-2 protein and / or the phosphoprotein P are suitable T-cell antigens and can promote an efficient T-cell response to the composition or vaccine when administered to a person.

[0075] The additional artificial RNA may comprise a coding sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to one of the SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11191, 21385-21388, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21411-21414 or a fragment or a This is a variant of one of these sequences.

[0076] Advantageously, the additional artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 9693-10052, 10143-10502, 10646-11005, 11192-11551, 19853-20212, 20222-20581, 20591-20950, 20960-21319, 21481-21488, 21627-21634, 21553-21560, 21699-21706 or a fragment or variant of any of these sequences.

[0077] Suitablely, the composition may comprise the mRNA, as defined in the claims, which is complexed, encapsulated or associated with one or more lipids, thereby forming lipid nanoparticles.

[0078] The composition may preferably comprise the artificial mRNA, as defined in the claims, which is complexed with one or more lipids, thereby forming lipid nanoparticles (LNPs), wherein the LNP essentially consists of (i) at least one cationic lipid as defined herein, preferably a lipid of formula (III), particularly preferably lipid III-3; (ii) a neutral lipid as defined herein, preferably 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC); (iii) a steroid or steroid analogue as defined herein, preferably cholesterol; and (iv) a PEG lipid as defined herein, e.g. PEG-DMG or PEG-cDMA, preferably a PEGylated lipid of formula (IVa); wherein (i) to (iv) are in a molar ratio of about 20-60% cationic lipid : 5-25% neutral lipid : 25-55% sterol ; 0.5-15% PEG lipid.

[0079] The present invention also relates to an RSV vaccine containing this artificial mRNA or this composition.

[0080] The present invention also relates to the first or second medical use of the artificial mRNA, the composition and the vaccine in the treatment or prophylaxis of an RSV infection.

[0081] In particular, the present invention is directed to the first or second medical use of the artificial mRNA, composition and vaccine, as defined in the claims, in the treatment or prophylaxis of an infection with RSV or a disease associated with such an infection.

[0082] The invention further relates to first and second medical uses of artificial RNA in the treatment or prevention of a disorder or disease in a patient, compositions, and vaccines. The invention further relates to a kit, in particular a kit of components, comprising the artificial RNA, compositions, and vaccines. Detailed description of the invention

[0083] The present application is filed together with a sequence protocol in electronic format, which forms part of the description of the present application (WIPO Standard ST.25). The information contained in the electronic version of the sequence protocol filed with the present application is incorporated herein by reference in its entirety. Reference here to a “SEQ ID NO” refers to the corresponding nucleic acid sequence or amino acid sequence (aa) in the sequence protocol with the respective identifier. For many sequences, the sequence protocol also contains additional detailed information, e.g., about specific structural features, sequence optimizations, GenBank identifiers, or additional detailed information about the coding capacity. In particular, such information is identified by the numerical identifier. <223> specified in the WIPO standard ST.25 sequence protocol.Accordingly, those under the numerical identifier <223> The information provided here is expressly included in its entirety and is to be understood as an integral part of the description of the underlying invention.

[0084] The following RSV fusion (F) protein amino acid and nucleic acid sequences (encoding an RSV fusion (F) protein) are not claimed, but serve merely for illustration: Artificial nucleic acid:

[0085] In a first aspect, the invention relates to an artificial mRNA in a composition comprising at least one artificial mRNA: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97% identical to SEQ ID NO: 4957, wherein the at least one antigenic peptide or protein is an F protein with a deleted C-terminus (F-del), and the RSV F protein includes a DSCav1 mutation (S155C, S290C, S190F and V207L), wherein the RSV F-protein comprises the two subunits F2 and F1 in a single polypeptide chain, F2 and F1 being linked via a linker element, where the F2 linker F1 protein aa104-aa144 is missing, wherein the RSV F protein includes at least one further mutation selected from A149C and Y458C, wherein the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO: 68..

[0086] In general, the RNA according to the invention can consist of a protein-coding region and 5' and / or 3' untranslated regions (UTRs). The 3' UTR is variable in sequence and size; it extends between the stop codon and the poly(A) tail. Importantly, the 3' UTR sequence contains several regulatory motifs that determine RNA turnover, stability, and localization, and thus many aspects of post-transcriptional regulation. In the medical application of RNA (e.g., immunotherapy, vaccination), the regulation of RNA translation into protein is of paramount importance for therapeutic safety and efficacy. The present inventors surprisingly discovered that certain combinations of 3' UTRs and / or 5' UTRs act synergistically to enhance the expression of surgically linked nucleic acid sequences encoding antigenic RSV peptides or proteins.Artificial RNA molecules containing the UTR combinations according to the invention advantageously enable the rapid and transient expression of high amounts of RSV antigenic peptides or proteins derived from RSV F. Accordingly, the artificial RNA provided here is particularly useful and suitable for various in vivo applications, including vaccination against RSV.

[0087] Advantageously, the artificial RNA can comprise at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR. In this context, a UTR of the invention comprises or consists of a nucleic acid sequence derived from a 5'-UTR or a 3'-UTR of any naturally occurring gene or a fragment, homolog, or variant thereof. Preferably, a 5'-UTR or a 3'-UTR of the invention is heterologous to the at least one coding sequence encoding the at least one antigenic peptide or protein derived from RSV F. Suitable heterologous 5'-UTRs or heterologous 3'-UTRs are derived from naturally occurring genes (not derived from RSV). In other embodiments, synthetically produced 5'-UTRs or 3'-UTRs may be used within the scope of the present invention.

[0088] In preferred embodiments, the at least one artificial RNA comprises at least one heterologous 3'-UTR.

[0089] Preferably, the at least one heterologous 3'-UTR comprises a nucleic acid sequence or consists of a nucleic acid sequence derived from a 3'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, most preferably a human gene, or from a variant of a 3'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, most preferably a human gene.

[0090] Preferably, the artificial RNA of the present invention comprises a 3'-UTR that can be derived from a gene relating to an RNA with an extended half-life (providing a stable RNA), e.g. a 3'-UTR as defined and described below.

[0091] Preferably, the at least one heterologous 3'-UTR comprises a nucleic acid sequence derived from a 3'-UTR of a gene, preferably encoding a stable mRNA, or from a homolog, fragment or variant of the gene.

[0092] In preferred embodiments of the first aspect, the artificial RNA of the invention comprises at least one heterologous 3'-UTR, wherein the at least one heterologous 3'-UTR comprises a nucleic acid sequence derived from a 3'-UTR of a gene selected from PSMB3, ALB7, alpha-globin (referred to as "muag"), CASP1, COX6B1, GNAS, NDUFA1 and RPS9, or from a homolog, fragment or variant of any of these genes.

[0093] ALB7-derived 3'-UTR: In preferred embodiments, the 3'-UTR comprises or consists of a nucleic acid sequence derived from the 3'-UTR of a vertebrate albumin gene or a variant thereof, preferably from the 3'-UTR of a mammalian albumin gene or a variant thereof, more preferably from the 3'-UTR of a human albumin gene or a variant thereof, more preferably from the 3'-UTR of the human albumin gene according to GenBank Accession Number NM_000477.5, or from a homolog, fragment or a variant thereof.Accordingly, the artificial RNA of the invention can comprise a 3' UTR derived from an ALB7 gene, wherein the 3' UTR derived from an ALB7 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 35 or 36 or a fragment or variant thereof.

[0094] Alpha-globin gene-derived 3'-UTR: In preferred embodiments, the 3'-UTR comprises or consists of a nucleic acid sequence derived from the 3'-UTR of a vertebrate alpha-globin gene (referred to as "muag") or a variant thereof, preferably from the 3'-UTR of a mammalian alpha-globin or a variant thereof, more preferably from the 3'-UTR of a human alpha-globin gene or a variant thereof, and more preferably from the 3'-UTR of the human alpha-globin gene. Accordingly, the RNA according to the invention can comprise a 3'-UTR derived from an alpha-globin gene, wherein the 3'-UTR derived from an alpha-globin gene comprises a nucleic acid sequence or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 37 or 38 or a fragment or variant thereof.

[0095] PSMB3-derived 3'-UTR: The artificial RNA of the invention may comprise a 3'-UTR derived from a 3'-UTR of a gene encoding a proteasome subunit beta type 3 protein (PSMB3) or a homolog, variant, fragment, or derivative thereof. Such 3'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3'-UTR of a proteasome subunit beta type 3 (PSMB3) gene, preferably from a vertebrate, more preferably from a mammal, most preferably from a human proteasome subunit beta type 3 (PSMB3) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferably encode a proteasome subunit beta type 3 (PSMB3) protein that corresponds to a human proteasome subunit beta type 3 (PSMB3) protein (UniProt Ref. No. P49720, entry version #183 dated August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 3'-UTR derived from a PSMB3 gene, wherein the 3'-UTR derived from a PSMB3 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 23 or 24 or a fragment or variant thereof.

[0096] CASP1-derived 3' UTR: The artificial RNA of the invention may comprise a 3' UTR derived from a 3' UTR of a gene encoding a caspase-1 (CASP1) protein or a homolog, variant, fragment, or derivative thereof. Such 3' UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3' UTR of a caspase-1 (CASP1) gene, preferably from a vertebrate, more preferably a mammalian, most preferably a human caspase-1 (CASP1) gene, or a homolog, variant, fragment, or derivative thereof.Accordingly, the RNA according to the invention can comprise a 3'-UTR derived from a CASP1 gene, wherein the 3'-UTR derived from a CASP1 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 25 or 26 or a fragment or variant thereof.

[0097] COX681-derived 3'-UTR: The artificial RNA of the invention may comprise a 3'-UTR derived from a 3'-UTR of a COX6B1 gene encoding a cytochrome c oxidase subunit 6B1 (COX6B1) protein, or a homolog, variant, fragment, or derivative thereof. Such 3'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3'-UTR of a cytochrome c oxidase subunit 6B1 (COX6B1) gene, preferably a vertebrate gene, more preferably a mammalian gene, and most preferably a human cytochrome c oxidase subunit 6B1 (COX6B1) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a cytochrome c oxidase subunit 6B1 (COX6B1) protein corresponding to a human cytochrome c oxidase subunit 6B1 (COX6B1) protein (UniProt Ref. No. P14854, entry version #166 dated August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 3'-UTR derived from a COX6B1 gene, wherein the 3'-UTR derived from a COX6B1 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 27 or 28 or a fragment or variant thereof.

[0098] GNAS-Derived 3'-UTR: The artificial RNA of the invention may comprise a 3'-UTR derived from a 3'-UTR of a GNAS gene encoding a guanine nucleotide-binding protein G(s) subunit alpha isoform (GNAS) protein, or a homolog, variant, fragment, or derivative thereof. Such 3'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3'-UTR of a guanine nucleotide-binding protein G(s) subunit alpha isoform (GNAS) gene, preferably from a vertebrate, more preferably from a mammalian guanine nucleotide-binding protein G(s) subunit alpha isoform (GNAS) gene, or a homolog, variant, fragment, or derivative thereof.The gene can preferably encode a guanine nucleotide-binding protein G(s) subunit alpha isoform (GNAS) protein corresponding to a human guanine nucleotide-binding protein G(s) subunit alpha isoform (GNAS) protein (UniProt Ref. No. P63092, entry version #153 of August 30, 2017). Accordingly, the artificial RNA of the invention may comprise a 3'-UTR derived from a GNAS gene, wherein the 3'-UTR derived from a GNAS gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 29 or 30 or a fragment or variant thereof.

[0099] NDUFA1-derived 3'-UTR: The artificial RNA according to the invention can comprise a 3'-UTR derived from a 3'-UTR of a gene encoding an NADH dehydrogenase [ubiquinone] 1-alpha subcomplex subunit 1 (NDUFA1) protein or a homolog, variant, fragment, or derivative thereof. Such 3'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3'-UTR of an NADH dehydrogenase [ubiquinone] 1-alpha subcomplex subunit 1 (NDUFA1) gene, preferably from a vertebrate, more preferably from a mammalian NADH dehydrogenase [ubiquinone] 1-alpha subcomplex subunit 1 (NDUFA1) gene, or a homolog, variant, fragment, or derivative thereof.The gene can preferentially encode an NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) protein corresponding to a human NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) protein (UniProt Ref. No. 015239, entry version #152 dated August 30, 2017). Accordingly, the artificial RNA of the invention may comprise a 3'-UTR derived from an NDUFA1 gene, wherein the 3'-UTR derived from an NDUFA1 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 31 or 32 or a fragment or variant thereof.

[0100] RPS9-derived 3'-UTR: The artificial RNA of the invention may comprise a 3'-UTR derived from a 3'-UTR of a gene encoding a 40S ribosome protein S9 (RPS9) or a homolog, variant, fragment, or derivative thereof. Such 3'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 3'-UTR of a 40S ribosome protein S9 (RPS9) gene, preferably from a vertebrate, more preferably from a mammal, most preferably from a human 40S ribosome protein S9 (RPS9) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferably encode a 40S ribosomal protein S9 (RPS9) protein that corresponds to a human 40S ribosomal protein S9 (RPS9) protein (UniProt Ref. No. P46781, Entry Version #179 of August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 3'-UTR derived from an RPS9 gene, wherein the 3'-UTR derived from an RPS9 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 33 or 34 or a fragment or variant thereof.

[0101] Further 3'-UTRs: In embodiments, the artificial RNA defined herein comprises a 3'-UTR as described in WO2016 / 107877. Particularly suitable 3'-UTRs are SEQ ID NOs: 1 to 24 and SEQ ID NOs: 49 to 318 from patent application WO2016 / 107877, or fragments or variants of these sequences. Accordingly, the 3'-UTRs of the artificial RNA of the present invention may comprise or consist of a corresponding RNA sequence of the nucleic acid sequence according to SEQ ID NOs: 1 to 24 and SEQ ID NOs: 49 to 318 of patent application WO2016 / 107877. In other embodiments, the artificial RNA defined herein comprises a 3'-UTR as described in WO2017 / 036580. Particularly suitable 3'-UTRs are the SEQ ID NOs: 152 to 204 from patent application WO2017 / 036580, or fragments or variants of these sequences.Accordingly, the 3'-UTR of the artificial RNA of the present invention may comprise or consist of a corresponding RNA sequence of the nucleic acid sequence according to SEQ ID NOs: 152 to 204 of patent application WO2017 / 036580.

[0102] In a preferred embodiment, the artificial RNA comprises at least one heterologous 5'-UTR.

[0103] In preferred embodiments, the at least one artificial nucleic acid defined herein, in particular the RNA defined herein, may comprise at least one heterologous 5'-UTR.

[0104] Preferably, the at least one 5'-UTR comprises a nucleic acid sequence or consists of a nucleic acid sequence derived from the 5'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, most preferably a human gene, or from a variant of the 5'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, most preferably a human gene.

[0105] Preferably, the artificial RNA of the present invention comprises a 5'-UTR that can be derived from a gene relating to an RNA with an extended half-life (providing a stable RNA), e.g. a 5'-UTR as defined and described below.

[0106] Preferably, the at least one heterologous 5'-UTR comprises a nucleic acid sequence derived from a 5'-UTR of a gene, preferably encoding a stable mRNA, or from a homolog, fragment or variant of the gene.

[0107] In preferred embodiments of the first aspect, the artificial RNA of the invention comprises at least one heterologous 5'-UTR, wherein the at least one heterologous 5'-UTR comprises a nucleic acid sequence derived from a 5'-UTR of a gene selected from HSD17B4, RPL32, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B and UBQLN2, or from a homolog, fragment or variant of one of these genes.

[0108] RPL32-derived 5' UTR: The artificial RNA of the invention may comprise a 5' UTR derived from a 5' UTR of a gene encoding a 60S ribosomal protein L32 or a homolog, variant, fragment, or derivative thereof, wherein the 5' UTR preferably lacks the 5' TOP motif. Such 5' UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5' UTR of a 60S ribosomal protein L32 (RPL32) gene, preferably from a vertebrate, more preferably from a mammal, most preferably from a human 60S ribosomal protein L32 (RPL32) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5' UTR preferably does not include the 5' TOP motif of the gene. The gene can preferentially encode a 60S ribosomal protein L32 (RPL32) that corresponds to a human 60S ribosomal protein L32 (RPL32) (UniProt Ref. No. P62899, Entry Version #138 of 30.August 2017). Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an RPL32 gene, wherein the 5'-UTR derived from an RPL32 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 21 or 22 or a fragment or variant thereof.

[0109] HSD17B4-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding 17-beta-hydroxysteroid dehydrogenase 4 or a homolog, variant, fragment or derivative thereof, preferably without the 5'TOP motif. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a 17-beta-hydroxysteroid dehydrogenase 4 gene (also known as peroxisomal multifunctional enzyme type 2), preferably from a vertebrate, more preferably from a mammal, most preferably a human 17-beta-hydroxysteroid dehydrogenase 4 (HSD17B4) gene, or a homolog, variant, fragment or derivative thereof, wherein the 5'-UTR preferably does not include the 5'TOP of the gene.The gene may preferably encode a 17-beta-hydroxysteroid dehydrogenase 4 protein corresponding to human 17-beta-hydroxysteroid dehydrogenase 4 (UniProt Ref. No. Q9BPX1, Entry Version #139 of August 30, 2017) or a homolog, variant, fragment or derivative thereof. Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an HSD17B4 gene, wherein the 5'-UTR derived from an HSD17B4 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 1 or 2 or a fragment or variant thereof.

[0110] ASAH1-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding acid ceramidase (ASAH1, "acid ceramidase") or a homolog, variant, fragment, or derivative thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of an acid ceramidase (ASAH1) gene, preferably from a vertebrate, more preferably mammalian, most preferably human acid ceramidase (ASAH1) gene, or a homolog, variant, fragment, or derivative thereof. The gene may preferably encode an acidic ceramidase (ASAH1) protein corresponding to human acidic ceramidase (ASAH1) (UniProt Ref. No. Q13510, Entry Version #177 of June 7, 2017), or a homolog, variant, fragment or derivative thereof.

[0111] Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an ASAH1 gene, wherein the 5'-UTR derived from an ASAH1 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 3 or 4 or a fragment or variant thereof.

[0112] ATP5A1-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding the mitochondrial ATP synthase subunit alpha (ATP5A1) or a homolog, variant, fragment or derivative thereof, wherein the 5'-UTR preferably lacks the 5'TOP motif. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a mitochondrial ATP synthase subunit alpha (ATP5A1) gene, preferably from a vertebrate, more preferably from a mammal, and most preferably from a human mitochondrial ATP synthase subunit alpha (ATP5A1) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably does not include the 5'-TOP of the gene.The gene may preferably encode a mitochondrial ATP synthase subunit alpha protein corresponding to the human acidic mitochondrial ATP synthase subunit alpha protein (UniProt Ref. No. P25705, entry version #208 of August 30, 2017) or a homolog, variant, fragment or derivative thereof. Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an ATP5A1 gene, wherein the 5'-UTR derived from an ATP5A1 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 5 or 6 or a fragment or variant thereof.

[0113] MP68-derived 5'-UTR: The artificial RNA according to the invention can comprise a 5'-UTR derived from a 5'-UTR of a gene encoding MP68, or a homolog, fragment, or variant thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a 6.8 kDa mitochondrial proteolipid (MP68) gene, preferably from a vertebrate, more preferably from a 6.8 kDa mitochondrial proteolipid (MP68) gene of a mammal, or from a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a 6.8 kDa mitochondrial proteolipid (MP68) protein that corresponds to a human 6.8 kDa mitochondrial proteolipid (MP68) protein (UniProt Ref. No. P56378, entry version #127 of 15 February 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an MP68 gene, wherein the 5'-UTR derived from an MP68 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 7 or 8 or a fragment or variant thereof.

[0114] NDUFA4-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a cytochrome c oxidase subunit (NDUFA4), or a homolog, fragment, or variant thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a cytochrome c oxidase subunit (NDUFA4) gene, preferably from a vertebrate, more preferably from a mammalian cytochrome c oxidase subunit (NDUFA4) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a cytochrome c oxidase subunit (NDUFA4) protein corresponding to a human cytochrome c oxidase subunit (NDUFA4) protein (UniProt Ref. No. O00483, entry version #149 dated August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an NDUFA4 gene, wherein the 5'-UTR derived from an NDUFA4 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 9 or 10 or a fragment or variant thereof.

[0115] NOSIP-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a nitric oxide synthase-interacting (NOSIP) protein or a homolog, variant, fragment, or derivative thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a nitric oxide synthase-interacting (NOSIP) gene, preferably from a vertebrate, more preferably from a mammal, most preferably from a human nitric oxide synthase-interacting (NOSIP) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a nitric oxide synthase-interacting protein (NOSIP) that corresponds to a human nitric oxide synthase-interacting protein (NOSIP) (UniProt Ref. No. Q9Y314, Entry Version #130 of June 7, 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from a NOSIP gene, wherein the 5'-UTR derived from a NOSIP gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 11 or 12 or a fragment or variant thereof.

[0116] RPL31-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a 60S ribosomal protein L31 or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably lacks the 5'-TOP motif. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a 60S ribosomal protein L31 (RPL31) gene, preferably from a vertebrate, more preferably from a mammalian 60S ribosomal protein L31 (RPL31) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably does not include the 5'-TOP of the gene. The gene can preferentially encode a 60S ribosomal protein L31 (RPL31) that corresponds to a human 60S ribosomal protein L31 (RPL31) (UniProt Ref. No. P62899, entry version #138 of August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an RPL31 gene, wherein the 5'-UTR derived from an RPL31 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 13 or 14 or a fragment or variant thereof.

[0117] SLC7A3-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a cationic amino acid transporter 3 (solute carrier family 7 member 3, SLC7A3) protein or a homolog, variant, fragment, or derivative thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a gene for the cationic amino acid transporter 3 (SLC7A3), preferably from a vertebrate, more preferably from a mammalian gene for the cationic amino acid transporter 3 (SLC7A3), or a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a protein of the cationic amino acid transporter 3 (SLC7A3) that corresponds to a human protein of the cationic amino acid transporter 3 (SLC7A3) (UniProt Ref. No. Q8WY07, entry version #139 of August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from an SLC7A3 gene, wherein the 5'-UTR derived from an SLC7A3 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 15 or 16 or a fragment or variant thereof.

[0118] TUBB4B-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a tubulin beta-4B chain protein (TUBB4B) or a homolog, variant, fragment, or derivative thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a gene encoding the beta-4B chain of tubulin (TUBB4B), preferably from a vertebrate, more preferably from a mammalian, and most preferably from a human gene encoding the beta-4B chain of tubulin (TUBB4B), or a homolog, variant, fragment, or derivative thereof. The gene can preferentially encode a tubulin beta-4B chain (TUBB4B) protein that is identical to the human tubulin beta-4B chain (TUBB4B) protein (UniProt Ref. No. Q8WY07, entry version #142 dated August 30, 2017).Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from a TUBB4B gene, wherein the 5'-UTR derived from a TUBB4B gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 17 or 18 or a fragment or variant thereof.

[0119] UBQLN2-derived 5'-UTR: The artificial RNA of the invention may comprise a 5'-UTR derived from a 5'-UTR of a gene encoding a ubiquilin-2 (UBQLN2) protein, or a homolog, variant, fragment, or derivative thereof. Such 5'-UTRs preferably comprise or consist of a nucleic acid sequence derived from the 5'-UTR of a ubiquilin-2 (UBQLN2) gene, preferably from a vertebrate, more preferably from a mammalian ubiquilin-2 (UBQLN2) gene, or a homolog, variant, fragment, or derivative thereof. The gene may preferably encode a ubiquilin-2 (UBQLN2) protein, as described in UniProt Ref. No. Q9UHD9, Entry Version #151 dated August 30, 2017.Accordingly, the artificial RNA of the invention may comprise a 5'-UTR derived from a UBQLN2 gene, wherein the 5'-UTR derived from a UBQLN2 gene comprises or consists of a nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 19 or 20 or a fragment or variant thereof.

[0120] Further 5'-UTRs: In embodiments, the artificial RNA defined herein comprises a 5'-UTR as described in WO2013 / 143700. Particularly preferred 5'-UTRs are nucleic acid sequences derived from SEQ ID NOs: 1-1363, SEQ ID NO: 1395, SEQ ID NO: 1421 and SEQ ID NO: 1422 of patent application WO2013 / 143700, or fragments or variants thereof. In this context, it is preferred that the 5'-UTR of the artificial RNA according to the present invention comprises or consists of a corresponding RNA sequence of the nucleic acid sequence according to SEQ ID NOs: 1-1363, SEQ ID NO: 1395, SEQ ID NO: 1421 and SEQ ID NO: 1422 of patent application WO2013 / 143700. In other embodiments, the artificial RNA of the invention comprises a 5'-UTR as described in WO2016 / 107877.

[0121] Particularly preferred 5'-UTRs are nucleic acid sequences according to SEQ ID NOs: 25 to 30 and SEQ ID NOs: 319 to 382 of patent application WO2016 / 107877, or fragments or variants of these sequences. In this context, it is particularly preferred that the 5'-UTR of the artificial RNA comprises or consists of a corresponding RNA sequence of the nucleic acid sequence according to SEQ ID NOs: 25 to 30 and SEQ ID NOs: 319 to 382 of patent application WO2016 / 107877. In other embodiments, the artificial RNA of the invention comprises a 5'-UTR as described in WO2017 / 036580. Particularly preferred 5'-UTRs are nucleic acid sequences according to SEQ ID NOs: 1 to 151 of patent application WO2017 / 036580, or fragments or variants of these sequences. In this context, it is particularly preferred that the 5'-UTR of the artificial RNA comprises or consists of a corresponding RNA sequence of the nucleic acid sequence according to SEQ ID NOs: 1 to 151 of patent application WO2017 / 036580.

[0122] The inventors found that certain combinations of at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR advantageously enhance the translation of the at least one coding sequence that is surgically linked to the 3'-UTR and / or 5'-UTR and encodes at least one antigenic peptide or protein derived from an RSV F protein in the target tissue (e.g., muscle, dermis).

[0123] Accordingly, it is preferred that the at least one heterologous 5'-UTR, as defined herein, and the at least one heterologous 3'-UTR, as defined herein, act synergistically to enhance the production (i.e., translation) of the antigenic peptide or protein from the artificial RNA of the invention. These advantageous combinations of 5'-UTR and 3'-UTR are described below. Each of the abbreviations introduced below, namely “a-1”, “a-2”, “a-3”, “a-4”, “a-5”, “b-1”, “b-2”, “b-3”, “b-4”, “b-5”, “c-1”, “c-2”, “c-3”, “c-4”, “c-5”, “d-1”, “d-2”, “d-3”, “d-4”, “d-5”, “e-1”, “e-2”, “e-3”, “e-4”, “e-5”, “e-6”, “f-1”, “f-2”, “f-3”, “f-4”, “f-5”, “g-1”, “g-2”, “g-3”, “g-4”, “g-5”, “h-1”, “h-2”, “h-3”, “h-4”, “h-5”, “i-1”, “i-2”, “i-3”, are used throughout the description of the present invention and represent an advantageous combination of 5'-UTR and / or 3'UTR of the invention.

[0124] Accordingly, in preferred embodiments of the first aspect, the artificial RNA of the invention comprises a-1. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or a-2. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or a-3. at least one 5' UTR derived from a 5' UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or a-4. at least one 5' UTR derived from a 5' UTR of a NOSIP gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or a-5. at least one 5' UTR derived from a 5' UTR of an MP68 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or b-1. at least one 5' UTR derived from a 5' UTR of a UBQLN2 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or b-2. at least one 5' UTR derived from a 5' UTR of an ASAH1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or b-3. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or b-4. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or b-5. at least one 5' UTR derived from a 5' UTR of a NOSIP gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or c-1. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or c-2. at least one 5' UTR derived from a 5' UTR of a NOSIP gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or c-3. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or c-4. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or c-5. at least one 5'-UTR derived from a 5'-UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3'-UTR derived from a 3'-UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, or d-1. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a PSMB3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or d-2. at least one 5' UTR derived from a 5' UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or d-3. at least one 5' UTR derived from a 5' UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a GNAS gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or d-4. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or d-5. at least one 5' UTR derived from a 5' UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-1. at least one 5' UTR derived from a 5' UTR of a TUBB4B gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-2. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-3. at least one 5' UTR derived from a 5' UTR of an MP68 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-4. at least one 5' UTR derived from a 5' UTR of a NOSIP gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-5. at least one 5' UTR derived from a 5' UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or e-6. at least one 5' UTR derived from a 5' UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or f-1. at least one 5' UTR derived from a 5' UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a GNAS gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or f-2. at least one 5' UTR derived from a 5' UTR of an ATP5A1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or f-3. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or f-4. at least one 5' UTR derived from a 5' UTR of an HSD17B4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a GNAS gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or f-5. at least one 5' UTR derived from a 5' UTR of an MP68 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or g-1. at least one 5' UTR derived from a 5' UTR of an MP68 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or g-2. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or g-3. at least one 5' UTR derived from a 5' UTR of an NDUFA4 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a GNAS gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or g-4. at least one 5' UTR derived from a 5' UTR of a NOSIP gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or g-5. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or h-1. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or h-2. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a GNAS gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or h-3. at least one 5' UTR derived from a 5' UTR of an RPL31 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of an NDUFA1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or h-4. at least one 5' UTR derived from a 5' UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a CASP1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or h-5. at least one 5' UTR derived from a 5' UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3' UTR derived from a 3' UTR of a COX6B1 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof; or i-1. at least one 5'-UTR derived from a 5'-UTR of an SLC7A3 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3'-UTR derived from a 3'-UTR of an RPS9 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof. i-2. at least one 5'-UTR derived from a 5'-UTR of an RPL32 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof, and at least one 3'-UTR derived from a 3'-UTR of an ALB7 gene or from a corresponding RNA sequence, homolog, fragment or variant thereof. i-3. at least one 3'-UTR derived from a 3'-UTR of an alpha-globin gene or from a corresponding RNA sequence, homolog, fragment or variant thereof.

[0125] Advantageously, the artificial RNA of the invention comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from an RSV F protein specified herein, which is operatively linked to a 3' UTR and a 5' UTR composed of a-1 (HSD17B4 / PSMB3), a-2 (Ndufa4 / PSMB3), a-3 (SLC7A3 / PSMB3), a-4 (NOSIP / PSMB3), a-5 (MP68 / PSMB3), b-1 (UBQLN2 / RPS9), b-2 (ASAH1 / RPS9), b-3 (HSD17B4 / RPS9), b-4 (HSD17B4 / CASP1), b-5 (NOSIP / COX6B1), c-1 (NDUFA4 / RPS9), c-2 (NOSIP / NDUFA1), c-3 (NDUFA4 / COX6B1), c-4 (NDUFA4 / NDUFA1), c-5 (ATP5A1 / PSMB3), d-1 (Rpl31 / PSMB3), d-2 (ATP5A1 / CASP1), d-3 (SLC7A3 / GNAS), d-4 (HSD17B4 / NDUFA1), d-5 (SIc7a3 / Ndufa1), e-1 (TUBB4B / RPS9), e-2 (RPL31 / RPS9), e-3 (MP68 / RPS9), e-4 (NOSIP / RPS9), e-5 (ATP5A1 / RPS9), e-6 (ATP5A1 / COX6B1), f-1 (ATP5A1 / GNAS), f-2 (ATP5A1 / NDUFA1), f-3 (HSD17B4 / COX6B1), f-4 (HSD17B4 / GNAS), f-5 (MP68 / COX6B1), g-1 (MP68 / NDUFA1), g-2 (NDUFA4 / CASP1), g-3 (NDUFA4 / GNAS), g-4 (NOSIP / CASP1), g-5 (RPL31 / CASP1),h-1 (RPL31 / COX6B1), h-2 (RPL31 / GNAS), h-3 (RPL31 / NDUFA1), h-4 (SIc7a3 / CASP1), h-5 (SLC7A3 / COX6B1), i-1 (SLC7A3 / RPS9), i-2 (RPL32 / ALB7), or i-3 (α-globin gene).

[0126] In particularly preferred embodiments of the first aspect, the artificial RNA of the invention comprises UTR elements according to a-1 (HSD17B4 / PSMB3), a-4 (NDUFA4 / PSMB3), c-1 (NDUFA4 / RPS9), e-4 (NOSIP / RPS9), g-2 (NDUFA4 / CASP1), i-2 (RPL32 / ALB7) or i-3 (alpha-globin (muag)).

[0127] In a particularly preferred embodiment of the first aspect, the artificial RNA of the invention comprises UTR elements according to a-1 (HSD17B4 / PSMB3).

[0128] The invention relates to an artificial RNA, preferably an RNA suitable for vaccination, comprising at least one heterologous 5'-UTR as defined above, and / or at least one heterologous 3'-UTR as defined above, and at least one coding sequence operatively linked to the 3'-UTR and / or 5'-UTR, wherein the coding sequence encodes at least one antigenic peptide or protein derived from a respiratory syncytial virus (“RSV”) or a fragment or variant thereof.

[0129] As used here, the term “respiratory syncytial virus” or the corresponding abbreviation “RSV” is not restricted to any particular virus strain, variant, serotype or isolate, etc., encompassing any respiratory syncytial virus of any origin.

[0130] According to various embodiments, the artificial RNA, preferably the coding sequence of the artificial RNA, comprises a nucleic acid sequence or consists of a nucleic acid sequence derived from viruses, with the NCBI taxonomy ID (“NCBI ID”) listed in List 1 below. List 1: RSV virus strains:

[0131] Human orthopneumovirus, HRSV (NCBI ID 11250); Human respiratory syncytial virus A, HRSV-A, Respiratory syncytial virus group A (NCBI ID 208893); Human respiratory syncytial virus A strain Long, Human respiratory syncytial virus (subgroup A / strain Long) (NCBI ID 11260); Human respiratory syncytial virus A2, Human respiratory syncytial virus (strain A2), HRSVA (NCBI ID 11259); Human respiratory syncytial virus (strain RSB1734), (NCBI ID 11253); Human respiratory syncytial virus (strain RSB5857) (NCBI ID 11254); Human respiratory syncytial virus (strain RSB6190), (NCBI ID 11255); Human respiratory syncytial virus (strain RSB6256), (NCBI ID 11256); Human respiratory syncytial virus (strain RSB642), (NCBI ID 11252); Human respiratory syncytial virus (strain RSB6614), (NCBI ID 11257); Human respiratory syncytial virus B, HRSV-B, respiratory syncytial virus group B, (NCBI ID 208895);Human Respiratory Syncytial Virus 9320 (NCBI ID 253182); Human Respiratory Syncytial Virus B1 (NCBI ID 79692); Human Respiratory Syncytial Virus (Subgroup B / Strain 18537), (NCBI ID 11251); Human Respiratory Syncytial Virus (Subgroup B / Strain 8 / 60), (NCBI ID 11258); Human Respiratory Syncytial Virus S2, (NCBI ID 410078); Human Respiratory Syncytial Virus Strain RSS-2, (NCBI ID 11261); Unclassified Human Respiratory Syncytial Virus, (NCBI ID 410233); Human Respiratory Syncytial Virus (Strain RSP112 / Sweden / 02-03), (NCBI ID 410237); Human respiratory syncytial virus (strain RSP120 / Sweden / 02-03), (NCBI-ID 410238); Human respiratory syncytial virus (strain RSP121 / Sweden / 02-03), (NCBI-ID 410239); Human respiratory syncytial virus (strain RSP122 / Sweden / 02-03), (NCBI-ID 410247); Human respiratory syncytial virus (strain RSP13 / Sweden / 02-03), (NCBI-ID 410241);Human respiratory syncytial virus (strain RSP140 / Sweden / 02-03), (NCBI-ID 410248); Human respiratory syncytial virus (strain RSP16 / Sweden / 02-03), (NCBI-ID 410242); Human respiratory syncytial virus (strain RSP171 / Sweden / 02-03), (NCBI-ID 410246); Human respiratory syncytial virus (strain RSP183 / Sweden / 02-03), (NCBI-ID 410249); Human respiratory syncytial virus (strain RSP191 / Sweden / 02-03), (NCBI-ID 410240); Human respiratory syncytial virus (strain RSP199 / Sweden / 02-03), (NCBI-ID 410250); Human respiratory syncytial virus (strain RSP212 / Sweden / 02-03), (NCBI-ID 410251); Human respiratory syncytial virus (strain RSP41 / Sweden / 02-03), (NCBI-ID 410234); Human respiratory syncytial virus (strain RSP45 / Sweden / 02-03), (NCBI-ID 410235); Human respiratory syncytial virus (strain RSP56 / Sweden / 02-03), (NCBI-ID 410243); Human respiratory syncytial virus (strain RSP58 / Sweden / 02-03), (NCBI-ID 410236);Human respiratory syncytial virus (strain RSP67 / Sweden / 02-03), (NCBI-ID 410244); Human respiratory syncytial virus (strain RSP94 / Sweden / 02-03) (NCBI-ID 410245); Respiratory syncytial virus isolate RSV Memphis-37, (strain Memphis-37) (NCBI-ID 12814).;

[0132] In preferred embodiments of the invention, the at least one antigenic peptide or protein is derived from a respiratory syncytial virus isolate RSV Memphis-37 (strain Memphis-37) (NCBI ID: 12814). Within the scope of the present invention, including the information contained in the ST25 sequence protocol, the abbreviation “HRSV(Memphis-37)” is used for the particularly preferred RSV virus.

[0133] In preferred embodiments of the invention, the at least one antigenic peptide or protein is derived from a human respiratory syncytial virus A2, human respiratory syncytial virus (strain A2) (NCBI ID: 11259). Within the scope of the present invention, including the information contained in the ST25 sequence protocol, the abbreviation "HRSV(A2)" is used for the particularly preferred RSV virus.

[0134] It is understood that the person skilled in the art may also use amino acid sequences and nucleic acid sequences derived from any of the RSV strains listed in List 1 to adapt the teaching of the present invention and to obtain RNA constructs, compositions and vaccines according to the invention.

[0135] In various embodiments, the at least one antigenic peptide or protein can be selected from the fusion protein (F), the glycoprotein G, the short hydrophobic protein SH, the matrix protein M, the nucleoprotein N, the large polymerase L, the M2-1 protein, the M2-2 protein, the phosphoprotein P, the non-structural protein NS1 or the non-structural protein NS2 of the respiratory syncytial virus (RSV) or a fragment, variant or derivative thereof.

[0136] In particularly preferred embodiments of the first aspect, the at least one antigenic peptide or protein is derived from an RSV fusion protein (F). In this context, the amino acid sequence of the at least one antigenic peptide or protein can be selected from any peptide or protein derived from the RSV fusion protein F or from a fragment, variant, or derivative thereof.

[0137] The RSV F protein is initially expressed (after infection of a host cell) as a single polypeptide precursor, referred to as the full-length fusion protein F (hereinafter referred to as "F0"). F0 forms a trimer in the endoplasmic reticulum and is processed at two conserved sites by a cellular / host-specific furin-like protease, generating the polypeptides F1, F2, and Pep27. The Pep27 polypeptide is excised and is not part of the mature F protein. The F2 polypeptide is generated from the N-terminal portion of the F0 precursor and is linked to the F1 polypeptide via two disulfide bonds. The F1 polypeptide is generated from the C-terminal portion of the F0 precursor and anchors the mature F protein to the membrane via a transmembrane domain connected to a cytoplasmic tail. Three F2-F1 heterodimer units (“protomers”) assemble to form a mature F protein.Initially, the mature F protein exists in a metastable form (hereinafter referred to as the "prefusion conformation"). After induction, it undergoes a dramatic and irreversible conformational change (hereinafter referred to as the "postfusion conformation"), which leads to the fusion of the viral and target cell membranes.

[0138] Accordingly, the artificial RNA of the first aspect, preferably the artificial RNA suitable for vaccination, encodes at least one antigenic peptide or protein derived from an RSV-F protein or a fragment or variant thereof.

[0139] In preferred embodiments, at least one antigenic peptide or protein derived from an RSV-F protein may be derived from one of the following amino acid sequences (NCBI Protein Accession Numbers) listed in List 2 below. List 2: NCBI Protein Accession Numbers of the RSV fusion proteins (F):

[0140] Accession number protein, AJF44801.1, AJF44759.1, AJF44661.1, AJF44602.1, 2207424A, AAB38520.1, AAB38517.1, AAB38519.1, AAB38518.1, AVQ93587.1, AVQ93599.1, AVQ93571.1, AVQ93568.1, AVQ93589.1, AVQ93597.1, AVQ93563.1, AVQ93594.1, AVQ93606.1, AVQ93601.1, AVQ93562.1, AVQ93561.1, AVQ93607.1, AVQ93588.1 AVQ93575.1, AVQ93468.1, AVQ93467.1, AVQ93590.1, AVQ93552.1, AVQ93556.1, AVQ93471.1, AVQ93458.1, AVQ93494.1, AVQ93470.1, AVQ93489.1, AVQ93542.1, AVQ93472.1, AVQ93514.1, AVQ93485.1, AVQ93533.1, AVQ93481.1, AVQ93546.1, AVQ93512.1, AVQ93554.1, AVQ93551.1, AVQ93547.1, AVQ93558.1, AVQ93461.1, AVQ93500.1, AVQ93426.1, AVQ93398.1, AVQ93401.1, AVQ93361.1, AVQ93408.1, AVQ93443.1, AVQ93429.1, AVQ93359.1, AVQ93365.1, AVQ93366.1, AVQ93402.1, AVQ93377.1, AVQ93412.1, AVQ93391.1, AVQ93457.1, AVQ93372.1, AVQ93455.1, AVQ93364.1, AVQ93378.1, AVQ93393.1, AVQ93362.1, AVQ93585.1, ART28504.1, AVQ93404.1, AOS49123.1, AOS48496.1, AMT78271.1, AHX57174.1, AHW81390.1, AHV81506.1, AFX60128.1, AFX60129.1, AEQ63389.1, ARB66328.1, ANZ80034.1, AMN91253.1, P03420.1, AlO08046.1, NP_056863.1, AFX60234.1, AFX60231.1, AFX60232.1, AFX60222.1, AFX60219.1, AFX60215.1, AFX60214.1, AFX60212.1, AFX60208.1, AFX60202.1, AFX60220.1, AFX60213.1, AFX60190.1, AFX60187.1, AFX60201.1, AFX60173.1, AFX60169.1, AFX60162.1, AFX60156.1, AFX60151.1, AFX60150.1, AFX60148.1, AFX60141.1, AFX60127.1, AFX60137.1, AFX60135.1, AFV46420.1, AFX60200.1, AFV46419.1, AFV46417.1, AFV46413.1, AFV46414.1, AFV46410.1, AFV46403.1, AFV46409.1, AFP99061.1, AFM95400.1, AFV46401.1, AFP99064.1, AFM95376.1, AFX60138.1, AFP99060.1, AFM95365.1, AFM55563.1, AFM55530.1, AFM55442.1, AFM55420.1, AFM55552.1, AFM55365.1, AFP99059.1, AFM95385.1, AFM55354.1, AFM55343.1, AFM55387.1, AFM55299.1, AFM55288.1, AFM55266.1, AFM55277.1, AFM55255.1, AFM55222.1, AFM55211.1, AFI25262.1, AFD34266.1, AFM55332.1, AFD34264.1, AFD34262.1, AFD34265.1, AFD34261.1, AFD34260.1, AFD34259.1, AEQ63641.1, AEQ63487.1, AEQ63520.1, AEQ63378.1, AEQ63367.1, 4CCF_A, AEQ63334.1, AEO45949.1, AEO45939.1, AEQ63312.1, AEQ63586.1, AEO45919.1, AEO45909.1, AEO45889.1, AEO45879.1, AEO45869.1, AEO45929.1, AEO45850.1, AEO45859.1, AEQ63444.1, AEO23054.1, AEO23052.1, AEO23051.1, AEC32087.1, ADZ95785.1, AEC32085.1, ADZ95784.1, ADZ95783.1, ADZ95782.1, ADZ95781.1, ADZ95779.1 ADZ95780.1, ADZ95777.1, ADZ95778.1, ADZ95776.1, ADZ95775.1, ACY68435.1, ACO83302.1, ABI35685.1, AFI25251.1, AAX23994.1, AAQ97026.1, AAR14266.1, AAQ97027.1, AAQ97030.1, AAQ97028.1, AAC57027.1, AAQ97029.1, AAQ97031.1, AAM68160.1, AAM44851.1, P11209.2, P13843.1, AAO72325.1, AAM68157.1 CAA26143.1, 1512372A, AAB86664.1, AAO72324.1, AAB82446.1, AAO72323.1, AAM68154.1, AAA47410.1, P12568.1, ARB07894.1, AGG39517.1, BBC54612.1, BBC54636.1, BBC54627.1, BBC54621.1, BBC54570.1, BBC54579.1, BBC54595.1, BBC54555.1, BBC54552.1, BBC54564.1, BBC54581.1, BBC54571.1, BBC54553.1, BBC54565.1, BBC54245.1 BBC54243.1, BBC54238.1, BBC54242.1, BBC54239.1, BBC54235.1, BBC54234.1, BBC54236.1, BBC54244.1, BBC54186.1, BBC54178.1, BBC54202.1, BBC54151.1, BBC54142.1, BBC54134.1, BBC54170.1, BBC54210.1, BBC54169.1, BBC54213.1, BBC54203.1, BBC54160.1, BBC54163.1, BBC54215.1, BBC54156.1, BBC54179.1, BBC54150.1, BBC54207.1, BBC54194.1, BBC54149.1, BBC54138.1, BBC54199.1, BBC54220.1, BBC54181.1, BBC54132.1, BBC54146.1, BBC54122.1, BBC54124.1 BBB35202.1, BBB35201.1, BBB35192.1, BBB35193.1, BBB35199.1, BBB35184.1, BBB35126.1, BBB35133.1, BBB35130.1, BBB35160.1, BBB35162.1, BBB35181.1, BBB35165.1, BBB35121.1, BBB35138.1, BBB35176.1, BBB35142.1, BBB35136.1, BBB35153.1, BBB35115.1, BBB35150.1, BBB35097.1, BBB35109.1, BBB35094.1, BBB35183.1, BBB35104.1, BBB35099.1, BBB35188.1, AKA45871.1, ASV65838.1, AGG39373.1, All22107.1, AGG39400.1, AGG39457.1, ARR29240.1, ARR29251.1, ARR29189.1, ARR29207.1, AUH15164.1, ATV81343.1, AUC68654.1, AUC68577.1, AUC68566.1, AUC68555.1, AUC68478.1, AUC68445.1, AUC68500.1, AUC68522.1, AUC68291.1, AUC68149.1, AUC68094.1, AMA67097.1, AMA66866.1, AMA66580.1, AIZ95750.1, AIZ95541.1, AIZ95519.1, AHY21419.1, AHY21331.1,AHY21165.1, AHY21143.1, AHY21132.1, AGG39478.1,ATV93506.1,ATV93509.1, AIZ95717.1, ATV81354.1, ART28317.1, BBA57890.1, BBA57901.1, ASZ70099.1, ART28361.1, AQX36844.1, ASK05520.1, ART28427.1, ART28339.1, ART28297.1, ART28328.1, ARQ15966.1, ARN61507.1, ARA15413.1, AGG39394.1, APW78845.1, APW78867.1, APW78900.1, APW78878.1, APW78779.1, APW78702.1, APW78713.1, APW78680.1, APW78658.1, APW78647.1, APW78636.1, AOS48870.1, APW78614.1, AMT78905.1, AMT77402.1, AOZ15479.1, AGN28484.1, AHX57240.1, AHX57031.1, AOS48980.1, AOS48848.1, AOS48815.1, AOS48738.1, AOS49068.1, AOS48727.1, AOS48716.1, AOS48683.1, AOS48551.1, AOS48485.1, AOS48518.1, AOS48441.1, AOS48397.1, AOS48375.1, AOS48353.1, AOS48287.1, AOS48320.1, AOS48265.1, AOS48254.1, AOS48221.1, ANZ79638.1, ALC74025.1, AHV81286.1, AOD40888.1, AOD40516.1, AOD40214.1, AOD40125.1, AOD40104.1, AOD40082.1, AOD39908.1, AJF44826.1, AJF44506.1, AJF44535.1, ANZ80463.1, ANZ80408.1, ANZ80397.1, ANZ80386.1, ANZ80331.1, ANZ80320.1, ANZ80364.1, ANZ80276.1, ANZ80221.1, ANZ80188.1, ANZ80144.1, ANZ80133.1, ANZ80111.1, ANZ80122.1, ANZ80056.1, ANZ80012.1, ANZ80067.1, ANZ79979.1, ANZ79935.1, ANZ79990.1, ANZ79902.1, ANZ79759.1, ANZ79715.1, ANZ79671.1, AMT79718.1, AMT79586.1, AMT79553.1, AMT79542.1 AMT79476.1, AMT79388.1, AMT79201.1, AMT79190.1, AMT79157.1, AMT79124.1, AMT79091.1, AMT79047.1, AMT79014.1, AMT79003.1, AMT78872.1, AMT78832.1, AMT78689.1, AMT78645.1, AMT78513.1, AMT78480.1, AMT78447.1, AMT78392.1, AMT78194.1, AMT78084.1, AMT78051.1, AMT77963.1, AMT77908.1, AMT77776.1 AMT77710.1, AMT77424.1, AMN91385.1, AMN91264.1, AMN91242.1, AHX57504.1, CUS01881.1, CUS01880.1, CUS01877.1, CUS01874.1, CUS01870.1, CUS01875.1, AMA67350.1, AMA67251.1, AMA67262.1, AMA67229.1, AMA67196.1, AMA67130.1, AMA67185.1, AMA67163.1, AMA67086.1, AMA67075.1, AMA66998.1, AMA66987.1, AMA66976.1, AMA66965.1, AMA66921.1, AMA66844.1, AMA66833.1, AMA66624.1, AMA66613.1, AMA66591.1, AMA66569.1, AMA66547.1, AMA66503.1, AMA66492.1, AMA66481.1, AMA66448.1, AMA66415.1, AMA66393.1, AMA66360.1, AGG39553.1, AGG39469.1, AJZ70144.1, AJZ70166.1, AJZ70155.1, AJZ70133.1, AJZ70067.1, AJZ70001.1, AJZ69990.1, AJZ69968.1, AJZ69946.1, AJZ69913.1, AJZ69880.1, AJZ69847.1, AJZ69869.1, AJZ69770.1, AJZ69748.1, AJZ69726.1 AJZ69682.1, AJZ69671.1, AJZ69704.1, AJZ69660.1, AJZ69715.1, AJZ69627.1, AJZ69616.1, AJZ69638.1, AJO16077.1, AJO16055.1, AKE31881.1, AKE31882.1, AKE31878.1, AJF44835.1, AJF44790.1, AJF44737.1, AJF44716.1, AJF44725.1, AJF44643.1, AJF44628.1, AJF44624.1, AJF44613.1 AJF44566.1, AJF44555.1, AJF44526.1, AJF44517.1, AKA45882.1, AKA45849.1, AHA83913.1, AGN28715.1, AHA83902.1, AHA83837.1, AHA83826.1, AHA83705.1, AHA83630.1, AGG39505.1, AIY70242.1, AIY70198.1, All30203.1, AHY21463.1, AHY21397.1, AHY21320.1, AHY21298.1, AHY21287.1, AHY21276.1, AHY21254.1 AHY21199.1, AHY21176.1, AHX57537.1, AHX57471.1, AHX57427.1, AHX57042.1, AIZ95981.1, AlZ95893.1, AIZ95871.1, AIZ95816.1, AIZ95772.1, AIZ95629.1, AIZ95673.1, AlZ95596.1, AIZ95585.1, AIZ95552.1, AEQ63553.2, AEQ63542.2, AEQ63575.1, AEQ63531.1, AEQ63498.1, AEQ63422.1, AEQ63411.1, AHX57570.1, AHX57152.1, AHX57064.1, AHX57009.1, AHX56987.1, AHV82100.1, AHV82001.1, AHV81891.1, AHV81880.1, AHV81836.1, AHV81682.1, AHV81649.1, AHV81484.1, AHV81462.1, AHV81385.1, AHV81363.1, AHV81330.1, AHV81253.1, AHV81154.1, AHV81122.1, AHV81089.1, AHV81012.1, AHV80957.1, AHV80880.1, AHV80869.1, AHV80836.1, AHV80803.1, AGG39559.1, AGG39562.1, AGG39556.1, AGG39550.1, AGG39547.1, AGG39544.1, AGG39541.1, AGG39529.1, AGG39526.1 AGG39523.1, AGG39514.1, AGG39502.1, AGG39499.1, AGG39496.1, AGG39493.1, AGG39484.1, AGG39490.1, AGG39487.1, AGG39475.1, AGG39472.1, AGG39466.1, AGG39463.1, AGG39454.1, AGG39442.1, AGG39439.1, AGG39436.1, AGG39415.1, AGG39403.1, AGG39397.1, AGG39391.1, AGG39379.1 AHL84194.1, AHA84012.1, AHJ60043.1, BAO49770.1, BAO49766.1, BAO49767.1, AHA84034.1, AHA84023.1, AHA83990.1, AHA83957.1, AHA83924.1, AHA83891.1, AHA83880.1, AHA83782.1, AHA83760.1, AHA83694.1, AGT75357.1, AGN92848.1, AGN28792.1, AGN28781.1, AGN28759.1, AGN28748.1, AGN28693.1, AGN28638.1, AGN28627.1, AGN28539.1, AGN28528.1, AGN28462.1, AGN28440.1, AGL96787.1, AGL96786.1, AGL96784.1, AAS93662.1, AAS93657.1, AAS93656.1, AAS93659.1, AAS93660.1, AAS93663.1, AAS93664.1, AAS93655.1, CUS01869.1, AHG54517.1, ASF87348.1, ASF87341.1, ASF87344.1, ASF87351.1, ASF87338.1, ASF87342.1, ASF87352.1, ASF87337.1, ASF87336.1, AEQ98756.1, AEQ98757.1, AEQ98755.1, AEQ98752.1, AEQ98753.1, AEQ98747.1, ASF87325.1, ASF87326.1, 5W23_A, 5EA3_F, 5UDD_A, 5EA8_F, AHG54458.1, AEN74947.1, AHG54485.1, AHG54477.1, AHG54451.1, AHG54463.1, AHG54445.1, AEO12131.1, AEN74945.1, AEN74944.1, ASF87335.1, AHG54515.1, AHA61605.1, AHV81660.1, AHG54441,1 or AIY60640.1.

[0141] In the context of the present invention, “RSV-F protein”, “RSV fusion protein (F)”, “RSV F” or “F” can be understood in the broadest sense and refers to F0 (F polypeptide precursor), F1, F2 and Pep27 polypeptides, F2-F1 heterodimers or the mature F protein (consisting of three F2-F1 heterodimers) or fragments and variants thereof. Accordingly, the term “peptide or protein derived from an RSV fusion protein (F)” refers to a peptide, protein, fragment or variant derived, for example, from F0 (F protein polypeptide precursor), F1, F2 and Pep27 polypeptides, F2-F1 heterodimers or the mature F protein. Furthermore, the term “peptide or protein derived from an RSV fusion protein (F)” refers to a peptide, protein, fragment, or variant that is derived from an “RSV-F protein” or an “RSV fusion protein (F)” as defined above and can be genetically modified to, for example,Certain protein elements (e.g., the cytoplasmic tail, the furin cleavage site, Pep27) are missing or, for example, contain additional elements (e.g., linker elements, heterologous signal peptides, etc.). The term “peptide or protein derived from an RSV fusion protein (F)” refers, for example, to a peptide, protein, fragment, or variant derived from F0, F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F_DSCav1_mut4, F_DSCav1_mut5, F_DSCav1_mut6, F_DSCav1_mut7, F_DSCav1_mut8, F_DSCav1_mut0, F-del_DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8, F-del_DSCav1_mut0 (see Table 1 for an explanation of the constructs). Particularly suitable F-protein variants that can be encoded by the RNA of the first aspect are specified below and listed in Table 1.

[0142] It should be noted that when referring to amino acid residues (aa) and their position in an RSV F protein, any numbering used herein – unless otherwise specified – refers to the position of the respective aa residue in a corresponding F0 precursor protein of HRSV(A2) (SEQ ID NO: 68) or a corresponding F0 precursor protein of HRSV(Memphis-37) (SEQ ID NO: 8937 or 11726), with position “1” corresponding to the first aa residue, i.e., the aa residue at the N-terminus of an HRSV(A2) F0 precursor protein or an HRSV(Memphis-37) F0 precursor protein.

[0143] In preferred embodiments, the at least one coding sequence of the RNA of the first aspect encodes at least one antigenic peptide or protein from the RSV F-protein, wherein the RSV F-protein is a full-length F-protein (F0) or an F-protein with a deleted C-terminus (F-del) or a fragment or variant thereof.

[0144] In this context, any full-length RSV F protein (precursor protein, designated "F0") can be used as a suitable antigen and can preferably be derived from any of the NCBI protein access numbers listed in List 2 or selected from one of the SEQ ID NOs: 68, 8279-8967, or 11726. In preferred embodiments of the invention, the full-length F protein (F0) of HRSV(A2) (SEQ ID NO: 68) is used, see Overview Table 1. In other preferred embodiments of the invention, the full-length F protein (F0) of HRSV(Memphis-37) (SEQ ID NO: 8937 or 11726) is expediently used, see Overview Table 1.

[0145] In this context, any RSV F with a deleted C-terminus (F-del) can be used as a suitable antigen, preferably derived from one of the NCBI protein access numbers listed in List 2 or selected from one of the SEQ ID NOs 483, 8968-9683, or 12095. An example of such a deletion mutant is the RSV-Fdel554-574 protein according to (Oomens et al. 2006. J. Virol. 80(21):10465-77), in which the aa residues 554-574 of the F0 protein are completely removed. Deletion of most of the cytoplasmic tail (aa 554-574) of F0 leads to an improvement in intracellular trafficking / cell surface transport in vitro and significantly increases the cell surface expression of RSV F. The increased cell surface presentation leads to improved B-cell recognition (in accordance with published data; see WO2015024668).In preferred embodiments of the invention, F-protein with a deleted C-terminus, referred to here as “F-del” (SEQ ID NO: 483, 9653 or 12095), is used, see overview table 1. Given the high structural preservation of the RSV-F protein between different RSV strains (see e.g. Hause et al., 2017, PLOS ONE 12(6): e0180623), the deletion of aa 554-574 is applicable to different RSV-F protein sequences of different RSV isolates.

[0146] In particularly preferred embodiments, the artificial RNA of the first aspect encodes at least one antigenic peptide or protein derived from the RSV-F protein, wherein the RSV-F protein is designed to stabilize the antigen in a prefusion conformation. A prefusion conformation is particularly advantageous in the context of an efficient RSV vaccine because several potential epitopes for neutralizing antibodies are accessible only in this protein conformation.

[0147] In several embodiments, the RSV-F protein contains one or more amino acid substitutions that stabilize the F protein in the prefusion conformation, e.g., substitutions that stabilize the membrane-distant portion of the F protein (including the N-terminal region of the F1 polypeptide) in the prefusion conformation. The amino acid substitution can, for example, introduce a non-natural disulfide bond or be a cavity-filling amino acid substitution.

[0148] Accordingly, a preferred RSV F protein in several embodiments contains S155C and S290C substitutions forming a non-natural disulfide bond that stabilizes the protein in a prefusion conformation, i.e., a conformation that binds specifically to one or more antibodies with prefusion specification, and / or has a suitable antigenic site present in the prefusion conformation but not in the postfusion conformation of the RSV F protein. In further embodiments, the recombinant RSV F protein may additionally have an F, L, W, Y, H, or M substitution at position 190, position 207, or at positions 190 and 207.

[0149] An example of an RSV F protein designed to stabilize the antigen in the prefusion conformation is the RSV F protein with a DSCav1 mutation (S155C, S290C, S190F, and V207L) or a fragment or variant thereof. Such RSV F DSCav1 proteins have been described in the prior art (WO2014160463).

[0150] Accordingly, in particularly preferred embodiments, the artificial RNA of the first aspect encodes at least one antigenic peptide or protein derived from the RSV F protein, wherein the RSV F protein comprises a DSCav1 mutation (S155C, S290C, S190F and V207L) or a fragment or variant thereof.

[0151] Within the scope of the invention, it is understood that each RSV F is mutated at positions S155C, S290C, S190F, and V207L ​​to stabilize the protein in the prefusion conformation, and that it can be used appropriately within the scope of the invention. Accordingly, any of the above-mentioned NCBI protein access numbers, or any protein selected from SEQ ID NOs: 68, 8279-8967, 483, 8968-9683, 12095, or 11726, or fragments or variants thereof, can be selected by a person skilled in the art to introduce such amino acid modifications at S155C, S290C, S190F, and V207L ​​to generate different RSV F DSCav1 proteins.

[0152] In preferred embodiments, the full-length RSV F protein (precursor protein, “F0”) of HRSV(A2) (SEQ ID NO: 68) is used to introduce the amino acid modifications S155C, S290C, S190F, and V207L, resulting in an amino acid sequence according to SEQ ID NO: 898. Such an RSV F protein is referred to as “FO_DSCav1” within the scope of the present invention (see Overview Table 1 (preferred RSV F protein antigen designs)).

[0153] In other preferred embodiments, the RSV F_del protein (deleted cytoplasmic tail (aa 554-574)) of HRSV(A2) (SEQ ID NO: 483) is used to introduce the amino acid modifications S155C, S290C, S190F, and V207L, resulting in an amino acid sequence according to SEQ ID NO: 1267. Such an RSV F protein is referred to as “F-del_DSCav1” within the scope of the present invention (see Overview Table 1 (preferred RSV F protein antigen designs)).

[0154] In preferred embodiments, the full-length RSV F protein (precursor protein, “F0”) of HRSV(Memphis-37) (SEQ ID NO: 8937 or 11726) is used to introduce amino acid modifications to S155C, S290C, S190F, and V207L, resulting in an amino acid sequence according to SEQ ID NO: 12464. Such an RSV F protein is referred to as “FO_DSCav1” within the scope of the present invention (see Overview Table 1 (preferred RSV F protein antigen designs)).

[0155] In other preferred embodiments, the RSV F_del protein (deleted cytoplasmic tail (aa 554-574)) of HRSV(Memphis-37) (SEQ ID NO: 9653 or 12095) is used to introduce the amino acid modifications S155C, S290C, S190F, and V207L, resulting in an amino acid sequence according to SEQ ID NO: 12833. Such an RSV F protein is referred to as “F-del_DSCav1” within the scope of the present invention (see Overview Table 1 (preferred RSV F protein antigen designs)).

[0156] In preferred embodiments, the at least one antigenic peptide or protein can be an artificially produced protein comprising the two subunits F1 and F2 of mature F as a single polypeptide chain, wherein F2 and F1 are preferably linked via a linker (GS). Examples of these artificially produced F2-linker-F1 fusion proteins (e.g., F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) have been described in the scientific literature (Joyce, M. Gordon, et al. “Iterative structure-based improvement of a fusionglycoprotein vaccine against RSV”. Nature structural & molecular biology 23.9 (2016): 811; WO2017172890). The F2-linker F1-RSV-F proteins lack aa104-144 (containing the furin cleavage site and Pep27) and include a linker element between the F2 and F1 polypeptides (e.g., GS linker). These F2-linker F1 proteins may exhibit improved stability and / or antigenic properties.

[0157] The RSV-F protein comprises the two subunits F2 and F1 fused to form a single polypeptide chain, wherein F2 and F1 are linked via a linker element, preferably a GS linker as specified here, thereby preferably generating stable F2-linker-F1 proteins.

[0158] The F2-linker-F1 fusion proteins, e.g. F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553), plus a DScav1 mutation as described above (S155C, S290C, S190F and V207L).

[0159] The RSV F protein additionally includes at least one further mutation selected from (A149C, Y458C). In particularly preferred embodiments, the RSV F protein may additionally comprise at least one further mutation selected from (S46G, A149C, S215P, Y458C, K465Q), (S46G, E92D, A149C, S215P, Y458C, K465Q), (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q), (N183GC, N428C), (Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q), (Q98C, Q361C, L95M, I221M, R429K), or (N183GC, N428C, S46G, N67I, E92D, S215P, K465Q) or a fragment or variant thereof.

[0160] In particularly preferred embodiments, the F2-linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) may additionally, preferably in addition to the DSCav1 mutation, comprise at least one mutation selected from S46G, A149C, S215P, Y458C, K465Q, here referred to as “mut1”, e.g., SEQ ID NOs: 1636, 2005, or 14678, 15047; or S46G, E92D, A149C, S215P, Y458C, K465Q, here referred to as “mut2”, e.g., SEQ ID NOs: 2374, 2743 or 15416, 15785; or S46G, N67I, E92D, A149C, S215P, Y458C, K465Q, referred to here as “mut3”, e.g., SEQ ID NOs: 3112, 3481 or 13202, 13571, or a fragment or variant thereof (see overview Table 1 (preferred RSV F protein antigen designs)).

[0161] In other embodiments, the F2 linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) additionally comprise, in addition to the DSCav1 mutation, at least one mutation consisting of A149C, Y458C, or N183GC, N428C, or Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, 1221M, R429K, K465Q or Q98C, Q361C, L95M, I221M, R429K, " or N183GC, N428C, S46G, N67I, E92D, S215P, K465Q, or a fragment or variant thereof.

[0162] Within the scope of the invention, it is understood that each RSV F is adapted such that the two subunits F1 and F2 are contained in a single polypeptide chain, wherein F2 and F1 are linked via a linker, e.g. a (GS) linker, to increase the stability of the protein, as explained for the mutations “mut1”, “mut2” and “mut3”, by deletion of aa104-144 of the F0 polypeptide chain (as explained above), by introduction of a linker element between F2 and F1, as explained above, and by introduction of suitable amino acid substitutions, as explained above.Accordingly, all of the above-mentioned NCBI protein access numbers (see List 2) or any protein consisting of SEQ ID NOs: 68, 8279-8967, 483, 8968-9683, 11726, 12095 or fragments or variants thereof can be adapted by a person skilled in the art to generate F2 linker-F1 fusion proteins as described herein, and can be further adapted by introducing (S46G, A149C, S215P, Y458C, K465Q), (S46G, E92D, A149C, S215P, Y458C, K465Q), (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q), (A149C, Y458C), (N183GC, N428C), (Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q), (Q98C, Q361C, L95M, I221M, R429K), or (N183GC, N428C, S46G, N67I, E92D, S215P, K465Q) aa-substitutions and / or a DSCav1 mutation. Furthermore, apart from using a GS linker as described above, the skilled person can of course choose between various known linker elements to obtain similar, equally suitable RSV F protein variants (e.g.,selected from SEQ ID NOs: 117-162 of patent application WO2017 / 172890 or fragments or variants thereof, or selected from SEQ ID NOs: 1509-1565 of patent application WO2017 / 081082, or fragments or variants thereof).

[0163] In preferred embodiments, F2-linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprise a DSCav-1 mutation as claimed, and additionally an amino acid substitution mut1 as defined above, here referred to as F-del_DSCav1_mut1. In preferred embodiments, F2-linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprise a DSCav-1 mutation as claimed, and additionally an amino acid substitution mut2 as defined above, here referred to as F-del_DSCav1_mut2. In preferred embodiments, F2-linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) a DSCav-1 mutation as claimed, and additionally an amino acid substitution mut3 as defined above, here referred to as F-del_DSCav1_mut3.In preferred embodiments, F2-linker F1 proteins (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprise a DSCav-1 mutation as claimed, and additionally an amino acid substitution mut4 as defined above, here referred to as F-del_DSCav1_mut4.

[0164] A detailed description of the claimed and other RSV-F proteins can be found in Table 1.

[0165] In Table 1, all references to amino acid residues (aa) and their position in an RSV-F protein refer to the position of the respective aa residue in a corresponding F0 precursor protein of HRSV(A2) (SEQ ID NO: 68) or HRSV(Memphis-37) (SEQ ID NO: 8937 or 11726). Furthermore, throughout the entire description of the invention, the abbreviations for suitable RSV-F protein antigen designs in Table 1 are used (e.g., “F0”, “F-del”, “F0_DSCav1”, “F-del_DSCav1”, “F_DSCav1_mut1”, “F-del_DSCav1_mut1”, “F_DSCav1_mut2”, “F-del_DSCav1_mut2”, “F_DSCav1_mut3”, “F-del_DSCav1_mut3”, “F_DSCav1_mut4”, “F-del_DSCav1_mut4”, “F_DSCav1_mut5”, “F-del_DSCav1_mut5”, “F_DSCav1_mut6”, “F-del_DSCav1_mut6”, “F_DSCav1_mut7”, “F-del_DSCav1_mut7”, “F_DSCav1_mut8”, “F-del_DSCav1_mut8”, “F_DSCav1_mut0”, “F-del_DSCav1_mut0”).Column A of Table 1 contains the protein SEQ ID numbers of the respective RSV-F protein antigen designs derived from HRSV(A2); column B of Table 1 contains the protein SEQ ID numbers of the respective RSV-F protein antigen designs derived from HRSV(Memphis-37). In particular, the description of the invention expressly includes the information contained in Section 1. <223> The identifier of the ST25 sequence protocol of the present application is specified. Table 1: Claimed and other RSV-F protein antigen designs Antigen name Description of the protein design A B F0 aa1-574,RSV F0 precursor in full length 68 8937,11726 F-del aa1-553,Deletion of aa 554-574 of the C-terminus 483 9653,12095 F0_DSCav1 aa1-574,aa-Substitutions: S155C, S290C, S190F and V207L 898 12464 F-del_DSCav1 aa1-553, Deletion of aa 554-574 of the C-terminus, aa-Substitutions: S155C, S290C, S190F and V207L 1267 12833 F_DSCav1_mut0 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Konstruktaa-Substitutions: S155C, S290C, S190F and V207L 3850 13940 F-del_DSCav1_mut0 aa1-103 - GS(Linker) - aa145-553Deletion of aa 554-574 of the C terminus,F2 linker F1 constructaa substitutions: S155C, S290C, S190F and V207L 4219 14309 F_DSCav1_mut1 aa1-103 - GS(Linker) - aa145-574,F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;S46G, A149C, S215P, Y458C, K465Q 1636 14678 F-del_DSCav1_mut1 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; S46G, A149C, S215P, Y458C, K465Q 2005 15047 F_DSCav1_mut2 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;S46G, E92D, A149C, S215P, Y458C, K465Q 2374 15416 F-del_DSCav1_mut2 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; S46G, E92D, A149C, S215P, Y458C, K465Q 2743 15785 F_DSCav1_mut3 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;S46G, N67I, E92D, A149C, S215P, Y458C, K465Q 3112 13202 F-del_DSCav1_mut3 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; S46G, N67I, E92D, A149C, S215P, Y458C, K465Q 3481 13571 F_DSCav1_mut4 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;A149C, Y458C 4588 16154 F-del_DSCav1_mut4 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; A149C, Y458C 4957 16523 F_DSCav1_mut5 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;N183GC, N428C 5326 16892 F-del_DSCav1_mut5 aa1-103 - GS(linker) - aa145-553Deletion of aa 554-574 of the C terminus,F2-linker-F1 construct,aa substitutions: S155C, S290C, S190F and V207L;N183GC, N428C 5695 17261 F_DSCav1_mut6 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M,R429K, K465Q 6064 17630 F-del_DSCav1_mut6 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q 6433 17999 F_DSCav1_mut7 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;Q98C, Q361C, L95M, I221M, R429K 6802 18368 F-del_DSCav1_mut7 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; Q98C, Q361C, L95M, I221M, R429K 7171 18737 F_DSCav1_mut8 aa1-103 - GS(Linker) - aa145-574F2-Linker-F1-Construct,aa-Substitutions: S155C, S290C, S190F and V207L;N183GC, N428C, S46G, N67I, E92D, S215P, K465Q 7540 19106 F-del_DSCav1_mut8 aa1-103 - GS(Linker) - aa145-553 Deletion of aa 554-574 of the C-terminus, F2-Linker-F1 construct, aa-substitutions: S155C, S290C, S190F and V207L; N183GC, N428C, S46G, N67I, E92D, S215P, K465Q 7909 19475

[0166] In particularly preferred embodiments, the artificial RNA according to the first aspect encodes at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the RSV-F protein is selected from F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4 (see, for example, Table 1).

[0167] Particularly preferred and advantageous within the scope of the invention are RSV F proteins selected from F-del_DSCav1, F-del_DSCav1_mut0, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8 or a fragment or a variant thereof (see, for example, Table 1).

[0168] In preferred embodiments, the artificial mRNA in the pharmaceutical composition comprises at least one coding sequence encoding at least one antigenic peptide or protein comprising or consisting of at least one amino acid sequence that is identical or at least 97%, 98%, or 99% identical to one of the SEQ ID NOs: 2005, 2743, 3481, 4957, 16523. Additional information on each of these claimed amino acid sequences encoding RSV proteins can also be obtained from the sequence protocol, in particular from the entries therein under the identifier <223> The details are as follows.

[0169] According to another preferred embodiment, the artificial RNA of the invention encodes at least one antigenic peptide or protein as claimed and can additionally encode at least one further heterologous peptide or protein element.

[0170] The at least one additional peptide or protein element can suitably promote the secretion of the encoded antigenic peptide or protein of the invention (e.g., via secretory signal sequences), the anchoring of the encoded antigenic peptide or protein of the invention in the plasma membrane (e.g., via transmembrane elements), the formation of antigen complexes (e.g., via multimerization domains), or the formation of virus-like particles (VLP-forming sequence). Furthermore, the artificial nucleic acid sequence according to the present invention can additionally encode peptide linker elements, self-cleaving peptides, immunological adjuvant sequences, or dendritic cell target sequences. Suitable multimerization domains can be selected from the list of amino acid sequences according to SEQ ID NOs: 1116-1167 of patent application WO2017 / 081082, or from fragments or variants of these sequences. Trimerization and tetramerization elements can be, for example,The fibritin foldon domain from Enterobacteriaceae phages T4, GCN4p1, GCN4p1, and p53 is selected from engineered leucine zippers (engineered α-helical coiled-coil peptides that assume a parallel trimeric state). In this context, the fibritin foldon domains of Enterobacteriaceae phages T4, GCN4p1, GCN4p1, and p53 are preferred. Suitable transmembrane elements can be selected from the list of amino acid sequences according to SEQ ID NOs: 1228-1343 of patent application WO2017 / 081082, or fragments or variants of these sequences. Suitable VLP-forming sequences can be selected from the list of amino acid sequences according to SEQ ID NOs: 1168-1227 of patent application WO2017 / 081082, or fragments or variants of these sequences. Suitable peptide linkers can be selected from the list of amino acid sequences according to SEQ ID NOs: 1509-1565 of patent application WO2017 / 081082, or fragments or variants of these sequences.Suitable self-cleaving peptides can be selected from the list of amino acid sequences according to SEQ ID NOs: 1434-1508 of patent application WO2017 / 081082, or fragments or variants of these sequences. Suitable immunological adjuvant sequences can be selected from the list of amino acid sequences according to SEQ ID NOs: 1360-1421 of patent application WO2017 / 081082, or fragments or variants of these sequences. Suitable target sequences for dendritic cells (DCs) can be selected from the list of amino acid sequences according to SEQ ID NOs: 1344-1359 of patent application WO2017 / 081082, or fragments or variants of these sequences. Suitable secretory signal peptides can be selected from the list of amino acid sequences according to SEQ ID NOs: 1-1115 and SEQ ID NO: 1728 of patent application WO2017 / 081082, or fragments or variants of these sequences. At the nucleic acid level, any nucleic acid sequence (e.g.,The RNA sequence (RNA sequence) that codes for such amino acid sequences is selected. The heterologous secretory signal sequence can increase the secretion of the encoded antigenic peptide or protein.

[0171] According to embodiments, the secretory signal sequence comprises an amino acid sequence that is identical to, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to, one of the SEQ ID NOs: 21329-21362 or a fragment or variant of one of these sequences. Additional information on each of these suitable amino acid sequences encoding secretory signal sequences can also be found in the sequence protocol, in particular the entries therein under the identifier <223> details provided.

[0172] It is understood that at the nucleic acid level, any RNA sequence encoding an amino acid sequence identical to SEQ ID NO: 2005, 2743, 3481, 4957, 16523, or any nucleic acid sequence (e.g., DNA sequence, RNA sequence) encoding amino acid sequences that are at least 97%, 98%, or 99% identical to one of the SEQ ID NO: 2005, 2743, 3481, 4957, 16523, can be selected and accordingly understood as a suitable coding sequence and can therefore be included in the artificial RNA of the first aspect of the invention.

[0173] In particularly preferred embodiments, the artificial RNA of the first aspect suitably comprises a coding sequence located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR.

[0174] In preferred embodiments, the artificial mRNA of the pharmaceutical composition comprises a coding sequence comprising at least one of the nucleic acid sequences that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, 21402 (see also Table 3-6). Additional information on each of these claimed nucleic acid sequences that encode can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0175] According to preferred embodiments, the artificial RNA is a modified and / or stabilized artificial RNA. The artificial RNA of the claimed pharmaceutical composition, or the modified and / or stabilized artificial RNA of the pharmaceutical composition, is an artificial mRNA or a modified and / or stabilized artificial mRNA, respectively.

[0176] According to preferred embodiments, the artificial RNA of the present invention can therefore be provided as a “stabilized artificial RNA”, i.e., as an RNA that exhibits improved resistance to degradation in vivo, and / or as an artificial RNA that exhibits improved stability in vivo, and / or as an artificial RNA that exhibits improved translatability in vivo. Specific suitable modifications for “stabilizing” the artificial RNA are described below.

[0177] Such stabilization can be achieved by providing a “dried RNA” and / or a “purified RNA,” as described herein. Alternatively or additionally, such stabilization can be achieved, for example, by a modified phosphate backbone of the artificial RNA of the present invention. A backbone modification in connection with the present invention is a modification in which phosphates of the backbone of the nucleotides contained in the RNA are chemically modified. Nucleotides that can preferably be used in this context contain, for example, a phosphorothioate-modified phosphate backbone, wherein preferably at least one of the phosphate oxygens contained in the phosphate backbone is replaced by a sulfur atom. The stabilized RNAs can also include, for example, nonionic phosphate analogs, such as…Alkyl and arylphosphonates, in which the charged phosphonate oxygen is replaced by an alkyl or aryl group, or phosphodiesters and alkylphosphorus triesters, in which the charged oxygen residue is in alkylated form. Such skeleton modifications typically include, without limitation, modifications from the group of methylphosphonates, phosphoramidates, and phosphorothioates (e.g., cytidine-5'-O-(1-thiophosphate)).

[0178] Suitable modifications that are able to “stabilize” the artificial RNA according to the invention are described below.

[0179] According to embodiments, the artificial mRNA of the claimed pharmaceutical composition is a modified artificial RNA, wherein the modification refers to chemical modifications that include both backbone modifications and sugar or base modifications.

[0180] In this context, a modified artificial RNA, as defined herein, may contain nucleotide analogs / modifications, e.g., backbone modifications, sugar modifications, or base modifications. A backbone modification in connection with the present invention is a modification in which phosphates of the nucleic acid backbone contained in a nucleic acid, e.g., an artificial RNA, are chemically modified. A sugar modification in connection with the present invention is a chemical modification of the sugar of the nucleotides of the RNA, as defined herein. Furthermore, a base modification in connection with the present invention is a chemical modification of the base portion of the nucleotides of the RNA. In this context, the nucleotide analogs or modifications are preferably selected from nucleotide analogs suitable for transcription and / or translation.

[0181] In particularly preferred embodiments of the present invention, the nucleotide analogues / modifications that can be incorporated into a modified nucleic acid or, in particular, into a modified RNA as described herein, are preferably selected from 2-amino-6-chloropurine riboside 5'-triphosphate, 2-aminopurine riboside 5'-triphosphate; 2-aminoadenosine-5'-triphosphate, 2'-amino-2'-deoxycytidine triphosphate, 2-thiocytidine-5'-triphosphate, 2-thiouridine-5'-triphosphate, 2'-fluorothymidine-5'-triphosphate, 2'-O-methyl-inosine-5'-triphosphate, 4-thiouridine-5'-triphosphate, 5-aminoallylcytidine-5'-triphosphate, 5-aminoallyluridine-5'-triphosphate, 5-bromocytidine-5'-triphosphate, 5-bromouridine-5'-triphosphate, 5-bromo-2'-deoxycytidine-5'-triphosphate, 5-bromo-2'-deoxyuridine-5'-triphosphate, 5-iodocytidine-5'-triphosphate, 5-iodo-2'-deoxycytidine-5'-triphosphate, 5-iodouridine-5'-triphosphate, 5-iodo-2'-deoxyuridine-5'-triphosphate, 5-methylcytidine-5'-triphosphate, 5-methyluridine-5'-triphosphate,5-Propynyl-2'-deoxycytidine-5'-triphosphate, 5-Propynyl-2'-deoxyuridine-5'-triphosphate, 6-Azacytidine-5'-triphosphate, 6-Azauridine-5'-triphosphate, 6-Chloropurineriboside-5'-triphosphate, 7-Deazaadenosine-5'-triphosphate, 7-deazaguanosine 5'-triphosphate, 8-azaadenosine 5'-triphosphate, 8-azidoadenosine 5'-triphosphate, benzimidazole riboside 5'-triphosphate, N1-methyl adenosine 5'-triphosphate, N1-methyl guanosine 5'-triphosphate, N6-methyl adenosine 5'-triphosphate, O6-methylguanosine 5'-triphosphate, pseudouridine 5'-triphosphate or Puromycin 5'-triphosphate, xanthosine 5'-triphosphate. Particularly preferred are nucleotides for base modifications selected from the group of base-modified nucleotides, consisting of 5-methylcytidine 5'-triphosphate, 7-deazaguanosine 5'-triphosphate, 5-bromcytidine 5'-triphosphate and pseudouridine 5'-triphosphate, pyridin-4-one ribonucleoside, 5-azauridine, 2-thio-5-azauridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine.5-Carboxymethyluridin, 1-Carboxymethylpseudouridin, 5-Propinyluridin, 1-Propinylpseudouridin, 5-Taurinomethyluridin, 1-Taurinomethylpseudouridin, 5-Taurinomethyl-2-thio-uridin, 1-Taurinomethyl-4-thio-uridin, 5-Methyl-uridin, 1-Methyl-Pseudouridin, 4-Thio-1-methyl-Pseudouridin, 2-Thio-1-methyl-Pseudouridin, 1-Methyl-1-deaza-Pseudouridin, 2-Thio-1-methyl-1-deaza-Pseudouridin, Dihydrouridin, Dihydropseudouridin, 2-Thio-Dihydrouridin, 2-Thio-Dihydropseudouridin, 2-Methoxyuridin, 2-Methoxy-4-thio-uridin, 4-Methoxy-pseudouridin und 4-Methoxy-2-thio-pseudouridin, 5-Aza-Cytidin, Pseudoisocytidin, 3-Methylcytidin, N4-Acetylcytidin, 5-Formylcytidin, N4-Methylcytidin, 5-Hydroxymethylcytidin, 1-Methyl-Pseudoisocytidin, Pyrrolo-Cytidin, Pyrrolo-Pseudoisocytidin, 2-Thio-Cytidin, 2-Thio-5-Methylcytidin, 4-Thio-pseudoisocytidin, 4-Thio-1-Methyl-Pseudoisocytidin, 4-Thio-1-methyl-1-deaza-pseudoisocytidin, 1-Methyl-1-Deaza-Pseudoisocytidin, Zebularin, 5-Aza-Zebularin, 5-Methyl-Zebularin,5-Aza-2-Thio-Zebularin, 2-Thio-Zebularin, 2-Methoxy-Cytidin, 2-Methoxy-5-Methyl-Cytidin, 4-Methoxy-Pseudoisocytidin und 4-Methoxy-1-Methyl-Pseudoisocytidin, 2-Aminopurin, 2,6-Diaminopurin, 7-Deaza-adenin, 7-Desaza-8-aza-adenin, 7-Desaza-2-aminopurin, 7-Deaza-8-aza-2-aminopurin, 7-Deaza-2,6-Diaminopurin, 7-Desaza-8-aza-2,6-Diaminopurin, 1-Methyladenosin, N6-Methyladenosin, N6-Isopentenyladenosin, N6-(cis-Hydroxyisopentenyl)adenosin, 2-Methylthio-N6-(cis-Hydroxyisopentenyl)adenosin, N6-Glycinylcarbamoyladenosin, N6-Threonylcarbamoyladenosin, 2-Methylthio-N6-Threonylcarbamoyladenosin, N6,N6-Dimethyladenosin, 7-Methyladenin, 2-Methylthioadenin und 2-Methoxyadenin, Inosin, 1-Methylinosin, Wyosin, Wybutosin, 7-Deaza-Guanosin, 7-Deaza-8-Aza-Guanosin, 6-Thio-Guanosin, 6-Thio-7-Deaza-Guanosin, 6-Thio-7-deaza-8-aza-guanosin, 7-Methyl-guanosin, 6-Thio-7-methyl-guanosin, 7-Methylinosin, 6-Methoxy-guanosin, 1-Methyl-guanosin, N2-Methyl-guanosin, N2,N2-Dimethyl-guanosin, 8-Oxo-guanosin,7-Methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine and N2,N2-dimethyl-6-thio-guanosine, 5'-O-(1-thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine, 5'-O-(1-thiophosphate)-guanosine, 5'-O-(1-thiophosphate)-uridine, 5'-O-(1-thiophosphate)-pseudouridine, 6-aza-cytidine, 2-thio-cytidine, alpha-thio-cytidine, pseudo-iso-cytidine, 5-aminoallyl-uridine, 5-iodo-uridine, N1-methyl-pseudouridine, 5,6-dihydrouridine, Alpha-thio-uridine, 4-thio-uridine, 6-aza-uridine, 5-hydroxy-uridine, deoxy-thymidine, 5-methyl-uridine, pyrrolo-cytidine, inosine, alpha-thio-guanosine, 6-methyl-guanosine, 5-methyl-cytdine, 8-oxo-guanosine, 7-deaza-guanosine, N1-methyl-adenosine, 2-amino-6-chloro-purine, N6-methyl-2-amino-purine, pseudo-isocytidine, 6-chloro-purine, N6-methyl-adenosine, alpha-thio-adenosine, 8-azido-adenosine, 7-deaza-adenosine. Particularly preferred and suitable within the scope of the invention are pseudouridine (ψ), N1-methylpseudouridine (m1ψ),5-Methylcytosine and 5-methoxyuridine. Accordingly, the artificial RNA defined here can comprise at least one modified nucleotide selected from pseudouridine (ψ), N1-methylpseudouridine (m1ψ), 5-methylcytosine, and 5-methoxyuridine.

[0182] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence, wherein the at least one coding sequence is a coding sequence modified with pseudouridine (ψ).

[0183] Accordingly, in preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition or the at least one coding sequence comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides.

[0184] In further preferred embodiments, the artificial RNA of the invention comprises at least one coding sequence, wherein the at least one coding sequence is an N1-methylpseudouridine (m1ψ)-modified coding sequence.

[0185] Accordingly, in preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition or the at least one coding sequence comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ) nucleotides.

[0186] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence, wherein the at least one coding sequence is a codon-modified coding sequence.

[0187] In preferred embodiments, the at least one coding sequence of the invention is a codon-modified coding sequence, wherein the amino acid sequence encoded by the at least one codon-modified coding sequence is preferably unmodified compared to the amino acid sequence encoded by the corresponding wild-type coding sequence.

[0188] The term “codon-modified coding sequence” refers to coding sequences that differ from the corresponding wild-type coding sequence in at least one codon (triplets of nucleotides that code for an amino acid). A codon-modified coding sequence can conveniently exhibit, within the scope of the invention, improved resistance to in vivo degradation and / or improved in vivo stability and / or improved in vivo translatability. Codon modifications, in the broadest sense, take advantage of the degeneracy of the genetic code, in which several codons code for the same amino acid and can be used interchangeably (see Table 2), to optimize / modify the coding sequence for in vivo applications, as described above.

[0189] In particularly preferred embodiments of the artificial mRNA of the claimed pharmaceutical composition, the at least one sequence is a codon-modified coding sequence, wherein the codon-modified coding sequence is selected from the C-maximized coding sequence, the CAI-maximized coding sequence, the coding sequence adapted to human codon use, the G / C content-modified coding sequence and the G / C-optimized coding sequence, or any combination thereof.

[0190] According to preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition can be modified, wherein the C content of the at least one coding sequence can be increased, preferably maximized, compared to the C content of the corresponding wild-type coding sequence (here referred to as the "C-maximized coding sequence"). The amino acid sequence encoded by the C-maximized coding sequence of the RNA is preferably unchanged compared to the amino acid sequence encoded by the corresponding coding sequence of the wild-type nucleic acid. The generation of a C-maximized nucleic acid sequence can advantageously be carried out using a modification method according to WO2015 / 062738. Throughout the entire disclosure of the invention, including the <223> -Identifier of the sequence protocol, C-maximized coding sequences of suitable RSV nucleic acid sequences are marked by the abbreviation “opt2”.

[0191] According to embodiments, the artificial mRNA of the claimed pharmaceutical composition can be modified, wherein the G / C content of the at least one coding sequence of the invention can be modified compared to the G / C content of the corresponding wild-type coding sequence (hereinafter referred to as the "G / C-content modified coding sequence"). In this context, the terms "G / C optimization" or "G / C content modification" refer to a nucleic acid, preferably an artificial nucleic acid of the invention, which comprises a modified, preferably an increased number of guanosine and / or cytosine nucleotides compared to the corresponding wild-type nucleic acid sequence. Such an increased number can be generated by substituting codons containing adenosine or thymidine nucleotides for codons containing guanosine or cytosine nucleotides.When an enriched G / C ratio occurs in a coding sequence of DNA or RNA, the degradation of the genetic code is exploited. Particularly in the case of RNA, sequences with an increased G (guanosine) / C (cytosine) ratio are more stable than sequences with an increased A (adenosine) / U (uracil) ratio. The amino acid sequence encoded by the G / C-modified nucleic acid sequence is preferably unmodified compared to the amino acid sequence encoded by the corresponding wild-type nucleic acid sequence. Preferably, the G / C content of the coding sequence of the artificial nucleic acid sequence, e.g., the RNA sequence of the present invention, is reduced by at least 10%, preferably by at least 20%, more preferably by at least 30%, most preferably by at least 40% compared to the G / C content of the coding sequence of the corresponding wild-type nucleic acid sequence (e.g.,RNA sequence) encoding an RSV antigen as defined herein, or a fragment or variant thereof.

[0192] According to preferred embodiments, the mRNA of the claimed pharmaceutical composition can be modified, wherein the G / C content of the at least one coding sequence of the invention can be optimized compared to the G / C content of the corresponding wild-type coding sequence (hereinafter referred to as the "G / C-optimized coding sequence"). "Optimized" in this context refers to a coding sequence in which the G / C content is preferably increased to the substantially highest possible G / C content. The amino acid sequence encoded by the G / C-optimized coding sequence of the nucleic acid sequence is preferably unchanged compared to the amino acid sequence encoded by the corresponding coding sequence of the wild-type nucleic acid.The generation of a G / C content-optimized nucleic acid sequence can conveniently be carried out using a G / C content optimization method according to WO2002 / 098443. Throughout the entire disclosure of the invention, including the <223> -Identifier of the sequence protocol, G / C-optimized coding sequences of suitable RSV nucleic acid sequences are indicated with the abbreviation “opt1, opt5, opt6, opt11”.

[0193] According to embodiments, the artificial mRNA of the claimed pharmaceutical composition can be modified, wherein the codons in the at least one coding sequence of the invention can be adapted to human codon usage (referred to here as the "coding sequence adapted to human codon usage"). Codons that code for the same amino acid occur with varying frequencies in a subject, e.g., a human. Accordingly, the coding sequence of the artificial RNA is preferably modified such that the frequency of the codons that code for the same amino acid corresponds to the naturally occurring frequency of that codon according to human codon usage, as shown, for example, in Table 2.For example, in the case of the amino acid Ala, the wild-type coding sequence is preferably adapted such that the codon "GCC" is used with a frequency of 0.40, the codon "GCT" with a frequency of 0.28, the codon "GCA" with a frequency of 0.22, and the codon "GCG" with a frequency of 0.10, etc. (see Table 2). Accordingly, such a method (as shown by way of example for Ala) is applied for each amino acid encoded by the coding sequence of the artificial nucleic acid according to the invention in order to obtain sequences adapted to human codon usage. Throughout the entire disclosure of the invention, including the <223> -Identifier of the sequence protocol, coding sequences of suitable RSV nucleic acid sequences adapted to human codon usage are marked by the abbreviation “opt3”. Table 2: Table of codon usage in humans with frequency data for each amino acid amino acid Codon frequency amino acid Codon frequency Ala GCG 0.10 Per CCG 0.11 Ala GCA 0.22 Per CCA 0.27 Ala GCT 0.28 Per CCT 0.29 Ala GCC* 0.40 Per CCC* 0.33 Cys TGT 0.42 gin CAG* 0.73 Cys TGC* 0.58 Gln CAA 0.27 Asp GAT 0.44 Arg AGG 0.22 Asp GAC* 0.56 Arg AGA* 0.21 Glu GAG* 0.59 Arg CGG 0.19 Glu GAA 0.41 Arg CGA 0.10 Phe TTT 0.43 Arg CGT 0.09 Phe TTC* 0.57 Arg CGC 0.19 Gly GGG 0.23 Ser AGT 0.14 Gly GGA 0.26 Ser AGC* 0.25 Gly GGT 0.18 Ser TCG 0.06 Gly GGC* 0.33 Ser TCA 0.15 His CAT 0.41 Ser TCT 0.18 His CAC* 0.59 Ser TCC 0.23 Island ATA 0.14 Thr ACG 0.12 lie ATT 0.35 Thr ACA 0.27 Island ATC* 0.52 Thr ACT 0.23 Lys AAG* 0.60 Thr ACC* 0.38 Lys AAA 0.40 Val GTG* 0.48 Leu TTG 0.12 Val GTA 0.10 Leu TTA 0.06 Val GTT 0.17 Leu CTG* 0.43 Val GTC 0.25 Leu CTA 0.07 Trp TGG* 1 Leu CTT 0.12 Tyr TAT 0.42 Leu CTC 0.20 Tyr TAC* 0.58 Met ATG* 1 Stopp TGA* 0.61 Asn AAT 0.44 Stopp TAG 0.17 Asn AAC* 0.56 Stopp TAA 0.22 *: häufigstes menschliches Codon

[0194] According to embodiments, the artificial RNA of the present invention can be modified, wherein the codon adaptation index (CAI) in the at least one coding sequence of the invention can be increased or preferably maximized (here referred to as the "CAl-maximized coding sequence"). Accordingly, it is preferred that all codons of the wild-type nucleic acid sequence that are relatively rare in, for example, a human cell are exchanged for a corresponding codon that is common in, for example, a human cell, wherein the common codon codes for the same amino acid as the relatively rare codon. Preferably, the most common codons are used for each encoded amino acid (see Table 2; the most common human codons are marked with asterisks).Advantageously, the artificial RNA according to the invention comprises at least one coding sequence, wherein the codon adaptation index (CAI) of the at least one coding sequence is at least 0.5, at least 0.8, at least 0.9, or at least 0.95. The codon adaptation index (CAI) of the at least one coding sequence is most preferred. In the case of the amino acid Ala, for example, the wild-type coding sequence is adapted such that the most common human codon, "GCC," is always used for this amino acid. Accordingly, such a method (as described for Ala) is applied for each amino acid encoded by the coding sequence of the artificial RNA according to the invention in order to obtain CAI-maximized coding sequences. Throughout the entire disclosure of the invention, including the <223> -Identification of the sequence protocol, CAl-maximized coding sequences of suitable RSV nucleic acid sequences are marked by the abbreviation “opt4”.

[0195] Accordingly, in a particularly preferred embodiment, the artificial mRNA of the pharmaceutical composition comprises at least one coding sequence comprising a codon-modified nucleic acid sequence that is identical to, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to, a codon-modified nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, 21402 or a fragment or variant of one of these sequences (see also Tables 3 and 4). Additional information on each of these suitable nucleic acid sequences that encode can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0196] In a particularly preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence comprising a codon-modified nucleic acid sequence that is identical or at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to one of the G / C-optimized or G / C-content-modified nucleic acid sequences according to the SEQ ID NOs: 2006, 2007, 2011, 2012, 2013, 2744, 2745, 2749, 2750, 2751, 3482, 3483, 3487, 3488, 3489, 4958, 4959, 4963, 4964, 4965, 16524, 16525, 16529, 16530, 16531, 21370, 21372, 21374, 21376, 21402 or a fragment or variant of any of these sequences (see also Tables 3 and 4; opt1, 5, 6, 11). Additional information on each of these suitable nucleic acid sequences that encode can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0197] In a preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence, which includes a codon-modified nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence adapted to human codon use according to the SEQ ID NOs: 2009, 2747, 3485, 4961, 16527, or a fragment or variant of one of these sequences (see also Tables 3 and 4; opt3). Additional information on each of these suitable coding nucleic acid sequences can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0198] In a particularly preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence, which includes a codon-modified nucleic acid sequence that is identical to, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to, one of the C-maximized nucleic acid sequences according to the SEQ ID NOs: 2008, 2746, 3484, 4960, 16526, or a fragment or variant of one of these sequences (see also Tables 3 and 4; opt2). Additional information on each of these suitable coding nucleic acid sequences can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0199] In a preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence, which includes a codon-modified nucleic acid sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to one of the CAI-maximized nucleic acid sequences according to SEQ ID NOs 2010, 2748, 3486, 4962, 16528, or a fragment or variant of one of these sequences (see also Tables 3 and 4; opt4). Additional information on each of these suitable coding nucleic acid sequences can also be found in the sequence protocol, in particular under the identifier <223> details provided.

[0200] In embodiments, the A / U ratio in the vicinity of the ribosome binding site of the artificial nucleic acid, in particular the artificial RNA of the invention, can be increased compared to the A / U ratio in the vicinity of the ribosome binding site of its respective wild-type nucleic acid. This modification (increased A / U ratio around the ribosome binding site) increases the efficiency of ribosome binding to the nucleic acid, preferably the RNA. Effective binding of the ribosomes to the ribosome binding site, in turn, results in efficient translation of the RNA. Accordingly, in a particularly preferred embodiment, the artificial nucleic acid according to the invention comprises a ribosome binding site, also referred to as a "Kozak sequence," which is identical to, or at least 80%, 85%, 90%, or 95% identical to, one of the sequences SEQ ID NOs: 41, 42, or fragments or variants thereof.

[0201] Preferred RSV polypeptide and nucleic acid coding sequences (“cds”) are listed in Tables 3A and 3B and 4A and 4B.

[0202] In Tables 3A and 3B, columns A to J represent specific suitable constructs of the invention derived from the RSV fusion protein (F), wherein column A provides suitable sequences for F0, column B provides suitable sequences for F-del, column C provides suitable sequences for F0_DSCav1, column D provides suitable sequences for F-del_DSCav1, column E provides suitable sequences for F_DSCav1_mut1, column F provides suitable sequences for F-del_DSCav1_mut1, column G provides suitable sequences for F_DSCav1_mut2, column H provides suitable sequences for F-del_DSCav1_mut2, column I provides suitable sequences for F_DSCav1_mut3, and column J provides suitable sequences for F_del_DSCav1_mut3. The SEQ ID NOs of the specific proteins, as indicated in the sequence protocol, are listed in row 2 (“PRT”). The SEQ ID NOs of the corresponding wild-type / unmodified coding sequences are given in line 3 (“wt”).The SEQ ID NOs of the corresponding codon-modified coding sequences for each protein construct are given in lines 4 to 10 (“opt1”, “opt2”, “opt3”, “opt4”, “opt5”, “opt6”, “opt11”). Coding sequences derived from HRSV(A2) are given in Table 3A, and coding sequences derived from HRSV(Memphis-37) are given in Table 3B. Further information is provided in the <223> -Identifier for each of the respective SEQ ID NO included in the sequence protocol. Table 3A: Claimed and other coding sequences encoding RSV F (columns AJ), derived from HRSV(A2) A B C D E F G H I J PRT 68 483 898 1267 1636 2005 2374 2743 3112 3481 wt 69 484 opt1 70, 71, 485, 899, 1268, 1637, 2006, 2375, 2744, 3113, 3482, 21363 486, 900, 1269, 1638, 2007, 2376, 2745, 3114, 3483, 21364 21365 21366 21369 21370 21371 21372 21373 21374 opt2 72 487 901 1270 1639 2008 2377 2746 3115 3484 opt3 73 488 902 1271 1640 2009 2378 2747 3116 3485 opt4 74 489 903 1272 1641 2010 2379 2748 3117 3486 opt5 75 490 904 1273 1642 2011 2380 2749 3118 3487 opt6 76 491 905 1274 1643 2012 2381 2750 3119 3488 opt11 77 492 906 1275 1644 2013 2382 2751 3120 3489 Table 3B: Claimed and other coding sequences for RSV F (columns AJ), derived from HRSV(Memphis-37) A B C D E F G H I J PRT 11726 12095 12464 12833 13940 14309 14678 15047 15416 15785 wt 11727 12096 opt1 11728, 12097, 12465, 12834, 13941, 14310, 14679, 15048, 15417, 15786, 21389 21390 12466, 12835, 13942, 14311, 14680, 15049, 15418, 15787, 21391 21392 21395 21396 21397 21398 21399 21400 opt2 11729 12098 12467 12836 13943 14312 14681 15050 15419 15788 opt3 11730 12099 12468 12837 13944 14313 14682 15051 15420 15789 opt4 11731 12100 12469 12838 13945 14314 14683 15052 15421 15790 opt5 11732 12101 12470 12839 13946 14315 14684 15053 15422 15791 opt6 11733 12102 12471 12840 13947 14316 14685 15054 15423 15792 opt11 11734 12103 12472 12841 13948 14317 14686 15055 15424 15793

[0203] In Tables 4A and 4B, columns K to V represent specific suitable constructs of the invention derived from the RSV fusion protein (F), wherein column K provides suitable sequences for F_DSCav1_mut0, column L provides suitable sequences for F-del_DSCav1_mut0, column M provides suitable sequences for F_DSCav1_mut4, column N provides suitable sequences for F_del_DSCav1_mut4, column O provides suitable sequences for F_DSCav1_mut5, column P provides suitable sequences for F_del_DSCav1_mut5, column Q provides suitable sequences for F_DSCav1_mut6, column R provides suitable sequences for F_del_DSCav1_mut6, column S provides suitable sequences for F_DSCav1_mut7, column T provides suitable sequences for F_del_DSCav1_mut7, and column U provides suitable sequences for Column V of F_DSCav1_mut8 provides suitable sequences for F-del_DSCav1_mut8. The SEQ ID NOs of the specific proteins, as indicated in the sequence protocol, are in line 2 (“PRT”).The SEQ ID NOs of the corresponding codon-modified coding sequences for each protein construct are given in lines 3 to 9 (“opt1”, “opt2”, “opt3”, “opt4”, “opt5”, “opt6”, “opt11”). Coding sequences derived from HRSV(A2) are given in Table 4A, and coding sequences derived from HRSV(Memphis-37) are given in Table 4B. Further information is provided in the <223> -Identifier for each of the respective SEQ ID NO included in the sequence protocol. Table 4A: Used and other coding sequences for RSV F (columns KV), derived from HRSV(A2) K L M N O p Q R S T U V PRT 3850 4219 4588 4957 5326 5695 6064 6433 6802 7171 7540 7909 opt1 3851, 4220, 4589, 4958, 5327, 5696, 6065, 6434, 6803, 7172, 7541, 7910, 3852, 4221, 4590, 4959, 5328, 5697, 6066, 6435, 6804, 7173, 7542, 7911, 21367 21368 21375 21376 21377 21378 21379 21380 21381 21382 21383 21384 opt2 3853 4222 4591 4960 5329 5698 6067 6436 6805 7174 7543 7912 opt3 3854 4223 4592 4961 5330 5699 6068 6437 6806 7175 7544 7913 opt4 3855 4224 4593 4962 5331 5700 6069 6438 6807 7176 7545 7914 opt5 3856 4225 4594 4963 5332 5701 6070 6439 6808 7177 7546 7915 opt6 3857 4226 4595 4964 5333 5702 6071 6440 6809 7178 7547 7916 opt11 3858 4227 4596 4965 5334 5703 6072 6441 6810 7179 7548 7917 Table 4B: Sought and other coding sequences encoding RSV F (column KV) derived from HRSV(Memphis-37) K L M N O P Q R S T U V PRT 13202 13571 16154 16523 16892 17261 17630 17999 18368 18737 19106 19475 opt1 13203, 13572, 16155, 16524, 16893, 17262, 17631, 18000, 18369, 18738, 19107, 19476, 13204, 13573, 16156, 16525, 16894, 17263, 17632, 18001, 18370, 18739, 19108, 19477, 21393 21394 21401 21402 21403 21404 21405 21406 21407 21408 21409 21410 opt2 13205 13574 16157 16526 16895 17264 17633 18002 18371 18740 19109 19478 opt3 13206 13575 16158 16527 16896 17265 17634 18003 18372 18741 19110 19479 opt4 13207 13576 16159 16528 16897 17266 17635 18004 18373 18742 19111 19480 opt5 13208 13577 16160 16529 16898 17267 17636 18005 18374 18743 19112 19481 opt6 13209 13578 16161 16530 16899 17268 17637 18006 18375 18744 19113 19482 opt11 13210 13579 16162 16531 16900 17269 17638 18007 18376 18745 19114 19483

[0204] In embodiments, the artificial mRNA of the claimed pharmaceutical composition is monocistronic, bicistronic or multicistronic.

[0205] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition is monocistronic.

[0206] The term "monocistronic nucleic acid" is recognized and understood by those skilled in the art and refers, for example, to an artificial RNA that comprises only one coding sequence, as defined here. The terms "bicistronic nucleic acid," "multicistronic nucleic acid," or "monocistronic RNA" used here are also recognized and understood by those skilled in the art and refer, for example, to an artificial RNA that can have two (bicistronic) or even more (multicistronic) coding sequences.

[0207] In embodiments, the artificial RNA of the invention is monocistronic, and the coding sequence of the monocistronic artificial RNA encodes at least two different antigenic peptides or proteins derived from RSV F, as defined herein, or a fragment or variant thereof. Accordingly, the at least one coding sequence of the monocistronic artificial RNA can encode at least two, three, four, five, six, seven, eight, or more antigenic peptides or proteins derived from an RSV, preferably an RSV F, as defined herein, linked with or without an amino acid linker sequence, wherein the linker sequence can comprise rigid linkers, flexible linkers, cleavable linkers (e.g., self-cleaving peptides), as defined above, or a combination thereof (referred to herein as "multi-antigen constructs / nucleic acid").

[0208] In embodiments, the artificial RNA of the invention is bicistronic or multicistronic and comprises at least two coding sequences, wherein the at least two coding sequences encode two or more different antigenic peptides or proteins derived from RSV, preferably RSV F, as defined herein, or a fragment or variant thereof. Accordingly, the coding sequences in a bicistronic or multicistronic artificial RNA suitably encode different antigenic proteins or peptides, as defined herein, or a fragment or variant thereof. Preferably, the coding sequences in the bicistronic or multicistronic artificial RNA can be separated by at least one IRES sequence (internal ribosomal entry site).Thus, the term "encoding two or more antigenic peptides or proteins" can, without restriction, mean that the bicistronic or multicistronic artificial RNA encodes, for example, at least two, three, four, five, six, or more (preferably different) antigenic peptides or proteins of different RSVs or their fragments or variants within the scope of the definitions given here. Alternatively, the bicistronic or multicistronic artificial RNA can encode, for example, at least two, three, four, five, six, or more (preferably different) antigenic peptides or proteins that originate from the same RSV or its fragments or variants within the scope of the definitions given here. In this context, suitable IRES sequences can be selected from the list of nucleic acid sequences according to SEQ ID NOs: 1566-1662 of patent application WO2017 / 081082, or fragments or variants of these sequences.

[0209] It is understood that, within the scope of the invention, certain combinations of coding sequences can be generated by any combination of monocistronic, bicistronic and multicistronic artificial nucleic acids and / or multi-antigen constructs / nucleic acids to obtain a nucleic acid composition that codes for several antigenic peptides or proteins, as defined herein.

[0210] Preferably, the artificial RNA, which comprises at least one coding sequence as defined herein, typically has a length of about 50 to about 20,000 or 500 to about 20,000 nucleotides or about 500 to about 20,000 nucleotides or about 500 to about 10,000 nucleotides or about 1,000 to about 10,000 nucleotides or preferably about 1,000 to about 5,000 nucleotides or more preferably about 1,000 to about 2,500 nucleotides.

[0211] According to a preferred embodiment, the artificial RNA of the first aspect can be an mRNA, a self-replicating RNA, a circular RNA or a replicon RNA.

[0212] In embodiments, the artificial RNA is a circular RNA. As used herein, "circular RNA" or "circRNAs" means a circular polynucleotide capable of encoding at least one antigenic peptide or protein, as defined herein. Accordingly, in preferred embodiments, the circular RNA comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV or a fragment or variant thereof, as defined herein. Circular RNAs can be produced by various methods known in the prior art. For example, US6210931 teaches a method for synthesizing circRNAs by inserting DNA fragments into a plasmid containing sequences capable of spontaneous cleavage and self-circularization.US5773244 teaches the production of circRNAs by constructing a DNA construct encoding an RNA cyclase ribozyme that expresses the DNA construct as RNA and allows subsequent self-splicing of the RNA, resulting in an intron-free circRNA in vitro. WO1992 / 001813 teaches a method for producing single-stranded circular nucleic acids by synthesizing a linear polynucleotide, combining the linear nucleotide with a complementary linking oligonucleotide under hybridization conditions, and ligating the linear polynucleotide. The skilled person may also use the methods described in WO2015 / 034925 or WO2016 / 011222 for the production of circular RNA.

[0213] In embodiments, the artificial mRNA of the claimed pharmaceutical composition is a replicon RNA. The term "replicon RNA" is recognized and understood by those skilled in the art and refers, for example, to optimized self-replicating artificial RNA constructs. Such constructs comprise replication elements (replicases) derived from alphaviruses and the substitution of the structural viral proteins by the artificial nucleic acid of interest (in the context of the invention, an artificial nucleic acid comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV). Alternatively, the replicase can be provided on an independent construct comprising a replicase RNA sequence derived, for example, from Semliki Forest virus (SFV), Sindbis virus (SIN), Venezuelan equine encephalitis virus (VEE), Ross River virus (RRV), or other viruses from the alphavirus family.Downstream of the replicase lies a subgenomic promoter that controls the replication of the artificial nucleic acid according to the invention, i.e., an artificial nucleic acid comprising at least one coding sequence that codes for at least one antigenic peptide or protein derived from RSV.

[0214] The artificial RNA of the claimed pharmaceutical composition is an mRNA.

[0215] The terms "RNA" and "mRNA" are recognized and understood by experts and refer, for example, to a ribonucleic acid molecule, i.e., a polymer consisting of nucleotides. These nucleotides are typically adenosine monophosphate, uridine monophosphate, guanosine monophosphate, and cytidine monophosphate monomers linked together by a so-called backbone. The backbone is formed by phosphodiester bonds between the sugar (ribose) of one monomer and the phosphate group of a second, adjacent monomer. The specific sequence of monomers is called the RNA sequence. The mRNA (messenger RNA) typically provides the nucleotide sequence that can be translated into the amino acid sequence of a specific peptide or protein.

[0216] The artificial mRNA of the claimed pharmaceutical composition can be produced by any method known in the field of technology, including chemical synthesis, such as solid-phase RNA synthesis, as well as in vitro methods, such as RNA in vitro transcription reactions.

[0217] In a preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition is obtained by RNA-in-vitro transcription.

[0218] Accordingly, the mRNA of the claimed pharmaceutical composition is preferably an in vitro transcribed mRNA.

[0219] The terms “RNA in vitro transcription” or “in vitro transcription” refer to a process in which RNA is synthesized in a cell-free system (in vitro). RNA can be obtained by DNA-dependent in vitro transcription of a suitable DNA template, which, according to the present invention, is a linearized plasmid DNA template or a PCR-amplified DNA template. The promoter for controlling the RNA in vitro transcription can be any promoter for any DNA-dependent RNA polymerase. Specific examples of DNA-dependent RNA polymerases are the T7, T3, SP6, or Syn5 RNA polymerases. In a preferred embodiment of the present invention, the DNA template is linearized with a suitable restriction enzyme before being subjected to RNA in vitro transcription.

[0220] Reagents used in RNA in vitro transcription typically include: a DNA template (linearized plasmid DNA or PCR product) with a promoter sequence that has a high binding affinity for its respective RNA polymerase, such as bacteriophage-encoded RNA polymerases (T7, T3, SP6, or Syn5); ribonucleotide triphosphates (NTPs) for the four bases (adenine, cytosine, guanine, and uracil); optionally, a cap analogue as defined herein (e.g., m7G(5')ppp(5')G (m7G, m7G(5')ppp(5')(2'OMeG)pG, or m7G(5')ppp(5')(2'OMeA)pG)); optionally, further modified nucleotides as defined herein; a DNA-dependent RNA polymerase that is able to bind to the promoter sequence within the DNA template (e.g.T7, T3, SP6 or Syn5 RNA polymerase); optionally a ribonuclease (RNase) inhibitor to inactivate any potentially contaminating RNase; optionally a pyrophosphatase to break down pyrophosphate, which may inhibit RNA in vitro transcription; MgCl2, the Mg. 2+ -ions as a cofactor for the polymerase; a buffer (TRIS or HEPES) to maintain a suitable pH, which may also contain antioxidants (e.g. DTT), and / or polyamines such as spermidine in optimal concentrations, e.g. a buffer system containing TRIS citrate, as disclosed in WO2017 / 109161.

[0221] In embodiments, the nucleotide mixture used in RNA in vitro transcription may additionally contain modified nucleotides, as defined herein. In this context, preferred modified nucleotides include pseudouridine (ψ), N1-methylpseudouridine (m1ψ), 5-methylcytosine, and / or 5-methoxyuridine.

[0222] In preferred embodiments, the nucleotide mixture (i.e., the proportion of each nucleotide in the mixture) used for RNA in vitro transcription reactions can be optimized for the given RNA sequence, preferably as described in WO2015 / 188933.

[0223] In embodiments where more than one different artificial RNA, as claimed as mRNA, needs to be produced, e.g., when 2, 3, 4, 5, 6, 7, 8, 9, 10 or more different artificial RNAs need to be produced (e.g., encoding different RSV F antigens or e.g., a combination of RSV F and RSV G; see second aspect), methods as described in WO2017 / 109134 may be used appropriately.

[0224] In connection with the production of RNA vaccines, it may be necessary to provide GMP-quality RNA. GMP-quality RNA can be produced according to a manufacturing process approved by regulatory authorities. Accordingly, in a particularly preferred embodiment, RNA production is carried out according to current Good Manufacturing Practice (GMP), incorporating various quality control steps at the DNA and RNA levels, preferably in accordance with WO2016 / 180430. In preferred embodiments, the RNA of the invention is GMP-quality RNA, in particular GMP-quality mRNA.

[0225] The obtained RNA products are preferably processed using PureMessenger. ® (CureVac, Tübingen, Germany; RP-HPLC according to WO2008 / 077592) and / or tangential flow filtration (as described in WO2016 / 193206).

[0226] In a further preferred embodiment, the RNA, in particular the purified mRNA of the claimed pharmaceutical composition, is freeze-dried according to WO2016 / 165831 or WO2011 / 069586 to obtain a temperature-stable, dried artificial RNA (powder) as defined herein. The RNA according to the invention, in particular the purified RNA, can also be dried by spray drying or spray freeze-drying according to WO2016 / 184575 or WO2016 / 184576 to obtain a temperature-stable RNA (powder) as defined herein.

[0227] Accordingly, the mRNA of the claimed pharmaceutical composition is a dried mRNA.

[0228] The term “dried (m)RNA” used here is to be understood as (m)RNA that has been lyophilized, spray-dried or spray freeze-dried as defined above to obtain a temperature-stable dried (m)RNA (powder).

[0229] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition is a purified mRNA.

[0230] The term “purified RNA” or “purified mRNA” used here refers to RNA that, after certain purification steps (e.g., HPLC, TFF, oligo d(T) purification, precipitation steps), has a higher purity than the starting material (e.g., in vitro transcribed RNA). Typical impurities that are substantially absent in purified RNA include peptides or proteins (e.g., enzymes derived from DNA-dependent RNA in vitro transcription, such as RNA polymerases, RNases, pyrophosphatase, restriction endonuclease, DNase), spermidine, BSA, truncated RNA sequences, RNA fragments (short double-stranded RNA fragments, truncated sequences, etc.), free nucleotides (modified nucleotides, conventional NTPs, cap analogs), DNA template / matrices fragments, buffer components (HEPES, TRIS, MgCl2), etc. Other potential impurities, such as...Impurities that may arise from fermentation processes include bacterial contaminants (bioburden, bacterial DNA) or contaminants originating from purification processes (organic solvents, etc.). Accordingly, it is desirable that the "RNA purity level" be as close as possible to 100%. For the RNA purity level, it is also desirable that the amount of full-length RNA transcripts be as close as possible to 100%. Accordingly, "purified RNA" as used here has a purity level of more than 75%, 80%, 85%, particularly 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and most preferably 99% or more. The purity level can be further specified by the following criteria: B. by analytical HPLC, where the percentages given above correspond to the ratio between the area of ​​the peak for the target RNA and the total area of ​​all peaks representing the byproducts. Alternatively, the purity level can be determined, e.g.,can be determined by analytical agarose gel electrophoresis or capillary gel electrophoresis.

[0231] It is understood that “dried (m)RNA,” as defined herein, and “purified (m)RNA,” as defined herein, or “GMP-grade mRNA,” as defined herein, may exhibit superior stability properties (in vitro, in vivo) and improved efficiency (e.g., better translatability of the mRNA in vivo) and are therefore particularly suitable within the scope of the invention. Furthermore, “dried (m)RNA,” as defined herein, and “purified (m)RNA,” as defined herein, or “GMP-grade mRNA” may be particularly suitable for medical use, as defined herein.

[0232] The artificial mRNA of the claimed pharmaceutical composition can be suitably modified by adding a 5' cap structure, which preferably stabilizes the nucleic acid as described herein. Accordingly, in preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises a 5' cap structure, preferably m7G, cap0 (e.g., m7G(5')ppp(5')G), cap1 (e.g., m7G(5')ppp(5')(2'OMeG) or m7G(5')ppp(5')(2'OMeA)), cap2, a modified cap0 structure, or a modified cap1 structure (generated using a cap analogue as defined below).

[0233] The term “5’-cap structure” used here is recognized and understood by those skilled in the art and refers, for example, to a modified nucleotide (cap analogue), in particular a guanine nucleotide, which is attached to the 5’ end of an RNA molecule, e.g., an mRNA molecule. Preferably, the 5’-cap is added via a 5’-5’ triphosphate bond (also called m7GpppN).Other examples of 5'-cap structures include glyceryl, an inverted deoxyabasic moiety, a 4',5'-methylene nucleotide, a 1-(beta-D-erythrofuranosyl) nucleotide, a 4'-thiononucleotide, a carbocyclic nucleotide, 1,5-anhydrohexitol nucleotide, L-nucleotides, an alpha nucleotide, a modified base nucleotide, a threo-pentofuranosyl nucleotide, an acyclic 3',4'-seco-nucleotide, an acyclic 3,4-dihydroxybutyl nucleotide, an acyclic 3,5-dihydroxypentyl nucleotide, a 3'-3'-inverted nucleotide moiety, a 3'-3'-inverted abasic moiety, and a 3'-2'-inverted nucleotide moiety. 3'-2'-inverted abasic moiety, 1,4-butanediol phosphate, 3'-phosphoramidate, hexyl phosphate, aminohexyl phosphate, 3'-phosphate, 3'-phosphorus thioate, phosphorodithioate or a bridging or non-bridging methylphosphonate unit.

[0234] Other 5'-cap structures that may be suitable within the scope of the present invention are cap1 (additional methylation of the ribose of the adjacent nucleotide of m7GpppN), cap2 (additional methylation of the ribose of the 2nd nucleotide downstream of m7GpppN), cap3 (additional methylation of the ribose of the 3rd nucleotide downstream of m7GpppN), cap4 (additional methylation of the ribose of the 4th nucleotide downstream of m7GpppN), ARCA (anti-reverse cap analog), modified ARCA (e.g., phosphothioate-modified ARCA), inosine, N1-methylguanosine, 2'-fluoroguanosine, 7-deazaguanosine, 8-oxoguanosine, 2-aminoguanosine, LNAguanosine, and 2-Azido-guanosine.

[0235] A 5' cap structure (cap0 or cap1) can also be formed during chemical RNA synthesis or, preferably, during RNA in vitro transcription (co-transcriptional capping) using cap analogues.

[0236] The term “cap analogue,” as used here, is recognized and understood by those skilled in the art and refers, for example, to a non-polymerizable dinucleotide that exhibits cap functionality by facilitating translation or localization and / or preventing the degradation of a nucleic acid molecule, particularly an RNA molecule, when incorporated at the 5' end of the nucleic acid molecule. Non-polymerizable means that the cap analogue is incorporated only at the 5' terminus because it lacks a 5' triphosphate and therefore cannot be elongated in the 3' direction by a template-dependent polymerase, particularly a template-dependent RNA polymerase. Examples of cap analogues include, but are not limited to, a chemical structure selected from the group consisting of m7GpppG, m7GpppA, and m7GpppC; unmethylated cap analogues (e.g., GpppG); dimethylated cap analogues (e.g. m2,7GpppG), trimethylated cap analogues (e.g.m2,2,7GpppG), dimethylated symmetrical cap analogues (e.g., m7Gpppm7G) or anti-reverse cap analogues (e.g., ARCA; m7,2'OmeGpppG, m7,2'dGpppG, m7,3'OmeGpppG, m7,3'dGpppG and their tetraphosphate derivatives). Other cap analogues have already been described (WO2008 / 016473, WO2008 / 157688, WO2009 / 149253, WO2011 / 015347, and WO2013 / 059475). Other suitable cap analogues in this context are described in WO2017 / 066793, WO2017 / 066781, WO2017 / 066791, WO2017 / 066789, WO2017 / 053297, WO2017 / 066782, WO2018075827 and WO2017 / 066797.

[0237] The 5'-cap structure can conveniently be added co-transcriptionally using cap analogues as defined here in an RNA in vitro transcription reaction as defined here.

[0238] In preferred embodiments, a modified cap1 structure is generated using a cap analogue as disclosed in WO2017 / 053297, WO2017 / 066793, WO2017 / 066781, WO2017 / 066791, WO2017 / 066789, WO2017 / 066782, WO2018075827 and WO2017 / 066797. In particular, all cap structures derivable from the structure disclosed in claims 1-5 of WO2017 / 053297 can be used appropriately to co-transcribe a modified cap1 structure. Furthermore, all cap structures that can be derived from the structure defined in claim 1 or claim 21 of WO2018075827 can be suitablely used to co-transcribe a modified cap1 structure.

[0239] Preferred cap analogues are the dinucleotide cap analogues m7G(5')ppp(5')G (m7G) or 3'-O-Me-m7G(5')ppp(5')G for the co-transcriptional generation of cap0 structures. Particularly preferred cap analogues are the trinucleotide cap analogues m7G(5')ppp(5')(2'OMeA)pG or m7G(51ppp(5')(2'OMeG)pG for the co-transcriptional generation of cap1 structures.

[0240] In this context, it is preferred that the artificial mRNA of the claimed pharmaceutical composition comprises a Cap1 structure, as defined above, which preferably leads to increased protein expression, e.g., through high capping efficiencies and increased translation efficiencies. Furthermore, the mRNA according to the invention, comprising a Cap1 structure, exhibits lower stimulation of the innate immune system compared to Cap0 constructs of the same nucleic acid sequence. Those skilled in the art know how to determine translation efficiencies, capping levels, and immune stimulation.

[0241] In a particularly preferred embodiment, the artificial mRNA of the claimed pharmaceutical composition of the invention comprises a cap1 structure, wherein the cap1 structure can be formed enzymatically or co-transcriptionally (e.g. using m7G(5')ppp(5')(2'OMeA)pG or m7G(5')ppp(5')(2'OMeG)pG analogues).

[0242] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises an m7G(5')ppp(5')(2'OMeA)pG cap structure. In such embodiments, the coding RNA comprises a 5'-terminal m7G cap and an additional methylation of the ribose of the adjacent nucleotide of m7GpppN, in this case a 2'O-methylated adenosine.

[0243] In other preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises an m7G(5')ppp(5')(2'OMeG)pG cap structure. In such embodiments, the coding mRNA comprises a 5'-terminal m7G cap and an additional methylation of the ribose of the adjacent nucleotide, in this case a 2'O-methylated guanosine.

[0244] Accordingly, whenever suitable RNA or mRNA sequences are referred to in connection with the invention, the first nucleotide of this RNA or mRNA sequence, i.e. the nucleotide downstream of the m7G(5')ppp structure, can be a 2'O-methylated guanosine or a 2'O-methylated adenosine.

[0245] Accordingly, in other embodiments, the artificial mRNA of the claimed pharmaceutical composition may comprise a 5' cap sequence element according to SEQ ID NOs: 43 or 21321, or a fragment or variant thereof. SEQ ID NO: 43: gggaga (RNA, mRNA 5' end); SEQ ID NO: 21321: aggaga (RNA; mRNA 5' end). The sequence information is inserted here because the nucleotide sequences do not meet the minimum sequence length requirement for the ST.26 XML sequence protocol (10 or more specific, defined nucleotides or 4 or more specific, defined amino acids (see Section 8 of WIPO STANDARD ST.26)).

[0246] In other embodiments, the 5' cap structure is added by enzymatic capping using capping enzymes (e.g., vaccinia virus capping enzymes, commercially available capping kits) to generate cap0, cap1, or cap2 structures. In other embodiments, the 5' cap structure (cap0, cap1) is added by enzymatic capping using immobilized capping enzymes, e.g., with a capping reactor (WO2016 / 193226).

[0247] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one poly(A) sequence, preferably comprising 30 to 150 adenosine nucleotides.

[0248] In preferred embodiments, the poly(A) sequence located at the 3' terminus comprises 10 to 500 adenosine nucleotides, 10 to 200 adenosine nucleotides, 40 to 200 adenosine nucleotides, or 40 to 150 adenosine nucleotides. In a particularly preferred embodiment, the poly(A) sequence comprises about 64 adenosine nucleotides. In further particularly preferred embodiments, the poly(A) sequence comprises about 75 adenosine nucleotides. In further particularly preferred embodiments, the poly(A) sequence comprises about 100 adenosine nucleotides.

[0249] The terms “poly(A) sequence”, “poly(A) tail”, or “3' poly(A) tail”, as used herein, are recognized and understood by those skilled in the art and denote, for example, a sequence of adenosine nucleotides typically located at the 3' end of an RNA molecule and comprising up to about 1000 adenosine nucleotides. Preferably, the poly(A) sequence is substantially homopolymeric, i.e., a poly(A) sequence of, for example, 100 adenosine nucleotides has a length of substantially 100 nucleotides. In other embodiments, the poly(A) sequence may be interrupted by at least one nucleotide that differs from an adenosine nucleotide; for example, a poly(A) sequence of, for example, 100 adenosine nucleotides may be interrupted by at least one nucleotide that differs from an adenosine nucleotide. B. 100 adenosine nucleotides have a length of more than 100 nucleotides (comprising 100 adenosine nucleotides and additionally the at least one nucleotide that differs from an adenosine nucleotide).

[0250] Within the scope of the present invention, a poly(A) sequence can be located within an mRNA or another nucleic acid molecule, such as in a DNA vector, for example in a vector that serves as a template for the production of an RNA, preferably an mRNA, e.g. by transcription of the DNA vector.

[0251] Preferably, the poly(A) sequence of the artificial mRNA is obtained during in vitro transcription of the RNA from a DNA template. In other embodiments, the poly(A) sequence is obtained in vitro by conventional methods of chemical synthesis without necessarily being transcribed from a DNA template. In other embodiments, poly(A) sequences are generated by enzymatic polyadenylation of the RNA (after in vitro RNA transcription) using commercially available polyadenylation kits and appropriate protocols known in the art, or alternatively by using immobilized poly(A) polymerases, e.g., using a polyadenylation reactor (as described in WO2016 / 174271).

[0252] In embodiments, the artificial mRNA of the claimed pharmaceutical composition may contain a poly(A) sequence derived from a vector and may include at least one additional poly(A) sequence generated by enzymatic polyadenylation, e.g. as described in WO2016 / 091391.

[0253] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one poly(C) sequence, preferably comprising 10 to 40 cytosine nucleotides.

[0254] In preferred embodiments, the poly(C) sequence located at the 3' terminus comprises 10 to 200 cytosine nucleotides, 10 to 100 cytosine nucleotides, 20 to 70 cytosine nucleotides, 20 to 60 cytosine nucleotides, or 10 to 40 cytosine nucleotides. In a particularly preferred embodiment, the poly(C) sequence comprises about 30 cytosine nucleotides.

[0255] The term “poly(C) sequence”, as used herein, is recognized and understood by those skilled in the art and is, for example, a sequence of cytosine nucleotides typically located at the 3' end of an RNA molecule and comprising up to about 200 cytosine nucleotides. Within the scope of the present invention, a poly(C) sequence may be located within an mRNA or another nucleic acid molecule, such as in a DNA vector, e.g., in a vector that serves as a template for the production of an RNA, preferably an mRNA, e.g., by transcription of the vector.

[0256] Preferably, the poly(C) sequence in the RNA sequence of the present invention is derived from a DNA template (DNA template) by in vitro RNA transcription. In other embodiments, the poly(C) sequence is obtained in vitro by conventional methods of chemical synthesis without necessarily being transcribed from a DNA template.

[0257] In other embodiments, the artificial mRNA of the claimed pharmaceutical composition does not comprise a poly(C) sequence as defined herein.

[0258] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one histone stem loop.

[0259] The term "histone stem loop," as used here, is recognized and understood by those skilled in the art and refers, for example, to nucleic acid sequences that occur predominantly in histone mRNAs. Examples of histone stem loop sequences are described in Lopez et al. (Davila Lopez et al. (2008), RNA, 14(1)). The stem loops in histone pre-mRNAs are typically followed by a purine-rich sequence, the so-called histone downstream element (HDE). These pre-mRNAs are processed in the nucleus by a single endonucleolytic cleavage approximately 5 nucleotides downstream of the stem loop, a reaction catalyzed by the U7 snRNP through base pairing of the U7 snRNA with the HDE.

[0260] Histone stem loop sequences / structures can advantageously be selected from histone stem loop sequences as disclosed in WO2012 / 019780, the disclosure relating to histone stem loop sequences / structures being incorporated herein by reference. A histone stem loop sequence that can be used within the scope of the present invention may preferably be derived from formulas (I) or (II) of patent application WO2012 / 019780. According to a further preferred embodiment, the RNA defined herein may comprise at least one histone stem loop sequence derived from at least one of the specific formulas (1a) or (IIa) of patent application WO2012 / 019780.

[0261] In a particularly preferred embodiment, the artificial RNA of the invention comprises at least one histone stem loop, wherein the histone stem loop comprises a nucleic acid sequence according to SEQ ID NO: 39 or 40 or a fragment or a variant thereof.

[0262] In other embodiments, the artificial RNA of the first aspect does not include a histone stem loop as defined here.

[0263] In further embodiments, the artificial RNA of the invention comprises a 3'-terminal sequence element. The 3'-terminal sequence element is understood to be a sequence element comprising a poly(A) sequence and a histone stem loop sequence, wherein the sequence element is arranged at the 3' terminus of the artificial RNA according to the invention.

[0264] In other embodiments, the artificial mRNA of the claimed pharmaceutical composition may comprise a 3'-terminal sequence element according to SEQ ID NOs: 44-63 or 21322-21328 or a fragment or variant thereof.

[0265] In preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence modified with pseudouridine (ψ).

[0266] Accordingly, in preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition or the at least one coding sequence comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides.

[0267] In further preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition comprises at least one coding sequence modified with N1-methylpseudouridine (m1ψ).

[0268] Accordingly, in preferred embodiments, the artificial mRNA of the claimed pharmaceutical composition or the at least one coding sequence comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ) nucleotides.

[0269] In preferred embodiments of the first aspect, the artificial RNA, preferably mRNA, preferably in the 5' to 3' direction comprises the following elements a) -i): a) 5'-cap structure, preferably as shown here; b) if applicable 5'-UTR as specified here, preferably at least one selected from SEQ ID NOs: 1-22; c) a ribosome binding site, preferably as specified here d) at least one coding sequence as claimed, preferably as specified in Table 3 and Table 4; e) 3'-UTR as specified here, preferably at least one selected from SEQ ID NOs: 23-38; f) optionally a poly(A) sequence, preferably as shown here; g) optionally a poly(C) sequence, preferably as shown here; h) optionally a histone trunk loop, preferably as shown here; i) optionally a 3'-terminal sequence element as specified herein, preferably according to SEQ ID NOs: 44-63 or 21322-21328; and wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0270] In further preferred embodiments of the first aspect, the artificial RNA, preferably mRNA, comprises the following elements, preferably in the 5' to 3' direction: a) 5'-cap structure, preferably as shown here, most preferably a Cap1 structure; b) a 3'-UTR element and a 5'-UTR element according to a-1, a-4, c-1, e-4, g-2, i-2 or i-3, as specified here; c) a ribosome binding site, preferably as specified here; d) at least one coding sequence as claimed, wherein the coding sequence is arranged between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, wherein the coding sequence is preferably selected from a sequence specified in Table 3 and Table 4; e) optionally a poly(A) sequence, preferably as specified here; f) optionally a poly(C) sequence, preferably as shown here; g) optionally a histone stem loop, preferably as shown here; h) optionally a 3'-terminal sequence element as specified here, preferably according to SEQ ID NOs: 44-63, 21322-21328, wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0271] In further preferred embodiments of the first aspect, the artificial RNA, preferably mRNA, comprises the following elements, preferably in the 5' to 3' direction: a) 5'-cap structure, preferably as shown here, most preferably a Cap1 structure; b) a 3'-UTR element and a 5'-UTR element according to a-1 or i-3, as specified here; c) a ribosome binding site, preferably as specified here d) at least one coding sequence according to the claims, wherein the coding sequence is arranged between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, wherein the coding sequence is preferably selected from one of the sequences specified in Table 3 and Table 4; e) optionally a histone trunk loop, preferably as shown here; f) a poly(A) sequence, preferably comprising about 100 adenosine nucleotides; g) optionally a 3'-terminal sequence element as specified here, preferably according to SEQ ID NOs: 21322-21328, wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0272] In further preferred embodiments of the artificial mRNA of the claimed pharmaceutical composition, the artificial mRNA comprises the following elements in the 5' to 3' direction: a) 5'-cap structure, preferably as shown here, most preferably a Cap1 structure; b) a 3'-UTR element and a 5'-UTR element according to a-1, a-4, c-1, e-4, g-2, i-2 or i-3, as specified here; c) a ribosome binding site, preferably as specified here; d) at least one coding sequence as claimed, wherein the coding region is located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, wherein the coding sequence is preferably selected from one of the SEQ ID NOs2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, 21402 (or fragments or variants thereof); e) a poly(A) sequence comprising approximately 64 adenosines; f) a poly(C) sequence comprising approximately 30 cysteines; g) a histone stem loop according to SEQ ID NO: 39 or 40, wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0273] In further preferred embodiments of the first aspect, the artificial RNA, preferably mRNA, comprises the following elements in the 5' to 3' direction: a) 5'-Cap structure as claimed, most preferred a Cap1 structure; b) a 3'-UTR element and a 5'-UTR element according to a-1, a-4, c-1, e-4, g-2, i-2 or i-3, as specified here; c) a ribosome binding site, preferably as specified here; d) at least one coding sequence as specified herein, wherein the coding region is located between the 5' UTR and the 3' UTR, preferably downstream of the 5' UTR and upstream of the 3' UTR, wherein the coding sequence is preferably selected from one of the SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, 21402 (or fragments or variants thereof); e) a poly(A) sequence comprising approximately 64 adenosines; f) a histone stem loop according to SEQ ID NO: 39 or 40, wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0274] Preferred RSV polypeptide, nucleic acid and mRNA sequences are listed in Table 5A, 5B and Table 6A, 6B.

[0275] In Tables 5A and 5B, the protein designs are listed in row 1. Columns A to J in these tables represent specific suitable constructs of the invention derived from the RSV fusion protein (F), with column A providing suitable sequences for F0, column B providing suitable sequences for F-del, column C providing suitable sequences for F0_DSCav1, column D providing suitable sequences for F-del_DSCav1, column E providing suitable sequences for F_DSCav1_mut1, column F providing suitable sequences for F-del_DSCav1_mut1, column G providing suitable sequences for F_DSCav1_mut2, column H providing suitable sequences for F_del_DSCav1_mut2, column l providing suitable sequences for F_DSCav1_mut3, and column J providing suitable sequences for F_del_DSCav1_mut3.

[0276] The protein designs are listed in row 1 (columns A to J), and the specific SEQ ID NOs of the proteins, as indicated in the sequence protocol, are listed in row 2 ("Protein", "PRT"). The SEQ ID NOs of the corresponding coding sequences for each protein construct are listed in row 3 ("cds", see Table 3 for different cds optimizations). Further information, e.g., about the type of codon-modified coding sequence (opt1, opt2, opt3, opt4, opt5, opt6, opt11, etc.), is provided in the <223> The SEQ ID NO identifier is given in the sequence protocol and in Table 3. The SEQ ID NOs of the corresponding mRNA constructs, comprising the coding sequences and suitable 3' and 5' UTRs according to the invention, are given in lines 4 to 47 (mRNA designs a-1 to i-3 as shown here). Table 5A lists mRNA sequences derived from HRSV(A2), and Table 5B lists mRNA sequences derived from HRSV(Memphis-37). Further information, e.g.,Information on the type of coding sequence (wt, opt1, opt2, opt3, opt4, opt5, opt6, opt11, etc.) contained in the mRNA constructs is available in the <223> -Identifier of the respective SEQ ID NO included in the sequence protocol. Table 5B: Claimed and other mRNA constructs encoding RSV F (column AJ) derived from HRSV(Memphis-37) 1 A B C D E F G H I J 2 Protein 11726 12095 12464 12833 13940 14309 14678 15047 15416 15785 3 Kodierende 11727- 12096- 12465- 12834- 13941- 14310- 14679- 15048- 15417- 15786- Sequenz 11734, 12103, 12472, 12841, 13948, 14317, 14686, 15055, 15424, 15793, 21389 21390 21391 21392 21395 21396 21397 21398 21399 21400 4 mRNA 11735- 12104- 12473- 12842- 13949- 14318- 14687- 15056- 15425- 15794- Design a-1 11742, 12111, 12480, 12849, 13956, 14325, 14694, 15063, 15432, 15801, 21489, 21491, 21493- 21496- 21505- 21508- 21511- 21514- 21517- 21520- 21490, 21492, 21495, 21498, 21507, 21510, 21513, 21516, 21519, 21522, 21635, 21637, 21639- 21642- 21651- 21654- 21657- 21660- 21663- 21666- 21636 21638 21641 21644 21653 21656 21659 21662 21665 21668 5 mRNADesign a-2 11743-11750 12112-12119 12481-12488 12850-12857 13957-13964 14326-14333 14695-14702 15064-15071 15433-15440 15802-15809 6 mRNADesign a-3 11751-11758 12120-12127 12489-12496 12858-12865 13965-13972 14334-14341 14703-14710 15072-15079 15441-15448 15810-15817 7 mRNADesign a-4 11759-11766 12128-12135 12497-12504 12866-12873 13973-13980 14342-14349 14711-14718 15080-15087 15449-15456 15818-15825 8 mRNADesign a-5 11767-11774 12136-12143 12505-12512 12874-12881 13981-13988 14350-14357 14719-14726 15088-15095 15457-15464 15826-15833 9 mRNADesign b-1 11775-11782 12144-12151 12513-12520 12882-12889 13989-13996 14358-14365 14727-14734 15096-15103 15465-15472 15834-15841 10 mRNADesign b-2 11783-11790 12152-12159 12521-12528 12890-12897 13997-14004 14366-14373 14735-14742 15104-15111 15473-15480 15842-15849 11 mRNADesign b-3 11791-11798 12160-12167 12529-12536 12898-12905 14005-14012 14374-14381 14743-14750 15112-15119 15481-15488 15850-15857 12 mRNADesign b-4 11799-11806 12168-12175 12537-12544 12906-12913 14013-14020 14382-14389 14751-14758 15120-15127 15489-15496 15858-15865 13 mRNADesign b-5 11807-11814 12176-12183 12545-12552 12914-12921 14021-14028 14390-14397 14759-14766 15128-15135 15497-15504 15866-15873 14 mRNADesign c-1 11815-11822 12184-12191 12553-12560 12922-12929 14029-14036 14398-14405 14767-14774 15136-15143 15505-15512 15874-15881 15 mRNADesign c-2 11823-11830 12192-12199 12561-12568 12930-12937 14037-14044 14406-14413 14775-14782 15144-15151 15513-15520 15882-15889 16 mRNADesign c-3 11831-11838 12200-12207 12569-12576 12938-12945 14045-14052 14414-14421 14783-14790 15152-15159 15521-15528 15890-15897 17 mRNADesign c-4 11839-11846 12208-12215 12577-12584 12946-12953 14053-14060 14422-14429 14791-14798 15160-15167 15529-15536 15898-15905 18 mRNADesign c-5 11847-11854 12216-12223 12585-12592 12954-12961 14061-14068 14430-14437 14799-14806 15168-15175 15537-15544 15906-15913 19 mRNADesign d-1 11855-11862 12224-12231 12593-12600 12962-12969 14069-14076 14438-14445 14807-14814 15176-15183 15545-15552 15914-15921 20 mRNADesign d-2 11863-11870 12232-12239 12601-12608 12970-12977 14077-14084 14446-14453 14815-14822 15184-15191 15553-15560 15922-15929 21 mRNADesign d-3 11871-11878 12240-12247 12609-12616 12978-12985 14085-14092 14454-14461 14823-14830 15192-15199 15561-15568 15930-15937 22 mRNADesign d-4 11879-11886 12248-12255 12617-12624 12986-12993 14093-14100 14462-14469 14831-14838 15200-15207 15569-15576 15938-15945 23 mRNADesign d-5 11887-11894 12256-12263 12625-12632 12994-13001 14101-14108 14470-14477 14839-14846 15208-15215 15577-15584 15946-15953 24 mRNADesign e-1 11895-11902 12264-12271 12633-12640 13002-13009 14109-14116 14478-14485 14847-14854 15216-15223 15585-15592 15954-15961 25 mRNADesign e-2 11903-11910 12272-12279 12641-12648 13010-13017 14117-14124 14486-14493 14855-14862 15224-15231 15593-15600 15962-15969 26 mRNADesign e-3 11911-11918 12280-12287 12649-12656 13018-13025 14125-14132 14494-14501 14863-14870 15232-15239 15601-15608 15970-15977 27 mRNADesign e-4 11919-11926 12288-12295 12657-12664 13026-13033 14133-14140 14502-14509 14871-14878 15240-15247 15609-15616 15978-15985 28 mRNADesign e-5 11927-11934 12296-12303 12665-12672 13034-13041 14141-14148 14510-14517 14879-14886 15248-15255 15617-15624 15986-15993 29 mRNADesign e-6 11935-11942 12304-12311 12673-12680 13042-13049 14149-14156 14518-14525 14887-14894 15256-15263 15625-15632 15994-16001 30 mRNADesign f-1 11943-11950 12312-12319 12681-12688 13050-13057 14157-14164 14526-14533 14895-14902 15264-15271 15633-15640 16002-16009 31 mRNADesign f-2 11951-11958 12320-12327 12689-12696 13058-13065 14165-14172 14534-14541 14903-14910 15272-15279 15641-15648 16010-16017 32 mRNADesign f-3 11959-11966 12328-12335 12697-12704 13066-13073 14173-14180 14542-14549 14911-14918 15280-15287 15649-15656 16018-16025 33 mRNADesign f-4 11967-11974 12336-12343 12705-12712 13074-13081 14181-14188 14550-14557 14919-14926 15288-15295 15657-15664 16026-16033 34 mRNADesign f-5 11975-11982 12344-12351 12713-12720 13082-13089 14189-14196 14558-14565 14927-14934 15296-15303 15665-15672 16034-16041 35 mRNADesign g-1 11983-11990 12352-12359 12721-12728 13090-13097 14197-14204 14566-14573 14935-14942 15304-15311 15673-15680 16042-16049 36 mRNADesign g-2 11991-11998 12360-12367 12729-12736 13098-13105 14205-14212 14574-14581 14943-14950 15312-15319 15681-15688 16050-16057 37 mRNADesign g-3 11999-12006 12368-12375 12737-12744 13106-13113 14213-14220 14582-14589 14951-14958 15320-15327 15689-15696 16058-16065 38 mRNADesign g-4 12007-12014 12376-12383 12745-12752 13114-13121 14221-14228 14590-14597 14959-14966 15328-15335 15697-15704 16066-16073 39 mRNADesign g-5 12015-12022 12384-12391 12753-12760 13122-13129 14229-14236 14598-14605 14967-14974 15336-15343 15705-15712 16074-16081 40 mRNADesign h-1 12023-12030 12392-12399 12761-12768 13130-13137 14237-14244 14606-14613 14975-14982 15344-15351 15713-15720 16082-16089 41 mRNADesign h-2 12031-12038 12400-12407 12769-12776 13138-13145 14245-14252 14614-14621 14983-14990 15352-15359 15721-15728 16090-16097 42 mRNADesign h-3 12039-12046 12408-12415 12777-12784 13146-13153 14253-14260 14622-14629 14991-14998 15360-15367 15729-15736 16098-16105 43 mRNADesign h-4 12047-12054 12416-12423 12785-12792 13154-13161 14261-14268 14630-14637 14999-15006 15368-15375 15737-15744 16106-16113 44 mRNADesign h-5 12055-12062 12424-12431 12793-12800 13162-13169 14269-14276 14638-14645 15007-15014 15376-15383 15745-15752 16114-16121 45 mRNADesign i-1 12063-12070 12432-12439 12801-12808 13170-13177 14277-14284 14646-14653 15015-15022 15384-15391 15753-15760 16122-16129 46 mRNADesign i-2 12071-12086 12440-12455 12809-12824 13178-13193 14285-14300 14654-14669 15023-15038 15392-15407 15761-15776 16130-16145 47 mRNADesign i-3 12087-12094 12456-12463 12825-12832 13194-13201 14301-14308 14670-14677 15039-15046 15408-15415 15777-15784 16146-16153

[0277] In Tables 6A and 6B, the protein designs are listed in row 1. In this document, columns K to V represent specific suitable constructs of the invention derived from the RSV fusion protein (F), wherein column K provides suitable sequences for F_DSCav1_mut0, column L provides suitable sequences for F-del_DSCav1_mut0, column M provides suitable sequences for F_DSCav1_mut4, column N provides suitable sequences for F_del_DSCav1_mut4, column O provides suitable sequences for F_DSCav1_mut5, column P provides suitable sequences for F_del_DSCav1_mut5, column Q provides suitable sequences for F_DSCav1_mut6, column R provides suitable sequences for F_del_DSCav1_mut6, column S provides suitable sequences for F_DSCav1_mut7, column T provides suitable sequences for F_del_DSCav1_mut7, and column U provides suitable sequences for F_DSCav1_mut8. Column V provides suitable sequences for F-del_DSCav1_mut8.

[0278] The protein designs are listed in the first row (columns K to V), and the specific protein SEQ ID NOs, as specified in the sequence protocol, are in row 2, “PRT”. The SEQ ID NOs of the corresponding coding sequences for each protein design are listed in row 3, “cds” (see Table 4 for various cds optimizations). Further information, e.g., about the type of codon-modified coding sequence (opt1, opt2, opt3, opt4, opt5, opt6, opt11, etc.), is in the <223> The SEQ ID NO identifier is included in the sequence protocol and in Table 4. The SEQ ID NOs of the corresponding mRNA constructs, comprising the coding sequences and suitable 3' and 5' UTRs according to the invention, are indicated in each subsequent row (mRNA constructs "a-1" to "i-3" as indicated here). Table 6A lists mRNA sequences derived from HRSV(A2), and Table 6B lists mRNA sequences derived from HRSV(Memphis-37).

[0279] Further information, e.g., about the type of coding sequence (wt, opt1, opt2, opt3, opt4, opt5, opt6, opt11, etc.) contained in the mRNA constructs, can be found in the <223> -Identifier of the respective SEQ ID NO included in the sequence protocol.

[0280] In preferred embodiments, the artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-2167 or a fragment or variant of any of these sequences, wherein optionally at least one or more than one or all uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0281] In preferred embodiments, the artificial RNA comprises (a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR) and (b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV F or a fragment or variant thereof, wherein - the artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence consisting of the group comprising SEQ ID NOs: 2014-2373, 21436-21438, 21582-21584 (encoding F-del_DSCav1_mut1) or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2752-3111, 21442-21444, 21588-21590 (encoding F-del_DSCav1_mut2) or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3490-3849, 21448-21450, 21594-21596 (encoding F-del_DSCav1_mut3) or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 4966-5325, 16532-16891, 21454-21456, 21600-21602, 21526-21528, 21672-21674 (encoding F-del_DSCav1_mut4) or a fragment or variant of any of these sequences; and wherein, if necessary, at least one or more than one or all of the uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0282] As outlined throughout the specification, additional information on suitable amino acid sequences or nucleic acid sequences (coding sequences, mRNA sequences) can also be derived from the sequence protocol, in particular from the information therein under the identifier <223> The information provided is explained below.

[0283] The numerical identifier <223> In the sequence protocol of SEQ ID NO: 68, for example, it reads as follows: “derived and / or modified (artificial) protein sequence from HRSV(A2)_F0”. It should be noted that throughout the sequence protocol, the sequences listed under the numerical identifier are… <223> The information provided follows the same structure: “<SEQUENCE_DESCRIPTOR> from<CONSTRUCT_IDENTIFIER> ". The<SEQUENCE_DESCRIPTOR> refers to the type of sequence (e.g., "derived and / or modified protein sequence", "derived and / or modified CDS (coding sequence)", "mRNA product design a-1 with derived and / or modified sequence", or "mRNA product design b-4 comprising a derived and / or modified sequence", or "mRNA product design c-5 comprising a derived and / or modified sequence", or "mRNA product design g-4 with derived and / or modified sequence", etc.) and whether the sequence includes or consists of a wild-type sequence (“wt”) or whether the sequence includes or consists of a sequence-optimized sequence (e.g., “opt1”, “opt2”, “opt3”, “opt4”, “opt5”, “opt6”, “opt11”; sequence optimizations are described in more detail below). The<SEQUENCE_DESCRIPTOR> , which is identified by the numerical identifier <223> The description provided by SEQ ID NO: 68 reads, for example, as follows: “derived and / or modified protein sequence (artificial)”.<CONSTRUCT_IDENTIFIER> , which is designated by the numerical identifier <223> The specified structure has the following forms: (“organism_construct name”, or “organism_accession number_construct name”) and is intended to help the person skilled in the art to explicitly derive suitable nucleic acid sequences (e.g., RNA, mRNA) that encode the same RSV protein according to the invention. For example, the<CONSTRUCT_IDENTIFIER> , which is identified by the numerical identifier <223> The SEQ ID NO: 68 is specified as follows: “HRSV(A2)_F0”.In this example, the corresponding protein sequence is derived from “HRSV(A2)” (organism), wherein the protein comprises the structural elements “F0” (construct name, F in full). If the person skilled in the art uses the construct identifier of SEQ ID NO: 68, namely “HRSV(A2)_F0”, they can easily obtain a list of suitable nucleic acid coding sequences, e.g., RNA-coding sequences and mRNA sequences, which can be readily retrieved from the sequence record of the present invention. Composition:

[0284] A second aspect relates to a composition which, as claimed, contains at least one artificial mRNA of the first aspect.

[0285] In a preferred embodiment of the second aspect, the composition comprises at least one artificial mRNA of the first aspect and optionally at least one pharmaceutically acceptable carrier.

[0286] The term "pharmaceutically acceptable carrier," as used herein, preferably includes the liquid or non-liquid base of the composition. If the composition is in liquid form, the carrier is preferably water, typically pyrogen-free water; isotonic saline solution; or buffered (aqueous) solutions, e.g., phosphate-, citrate-, etc.-buffered solutions. Water or, preferably, a buffer, particularly preferably an aqueous buffer, may be used, containing a sodium salt, preferably at least 50 mM of a sodium salt, a calcium salt, preferably at least 0.01 mM of a calcium salt, and optionally a potassium salt, preferably at least 3 mM of a potassium salt. In a preferred embodiment, the sodium, calcium, and optionally potassium salts may be in the form of their halides, e.g., chlorides, iodides, or bromides, or in the form of their hydroxides, carbonates, hydrogen carbonates, or sulfates, etc.Without being limited to these, examples of sodium salts include NaCl, Nal, NaBr, Na₂CO₃, NaHCO₃, and Na₂SO₄. Examples of optional potassium salts include KCl, KI, KBr, K₂CO₃, KHCO₃, and K₂SO₄. Examples of calcium salts include CaCl₂, Cal₂, CaBr₂, CaCO₃, CaSO₄, and Ca(OH)₂. Furthermore, organic anions of the aforementioned cations may be included in the buffer.

[0287] In embodiments, the composition defined herein may comprise a plurality or at least more than one of the artificial mRNAs as defined in connection with the first aspect or the second aspect of the invention.

[0288] In embodiments, the at least one mRNA contained in the composition is a bi- or multicistronic nucleic acid, in particular a bi- or multicistronic nucleic acid as defined herein, encoding at least two, three, four, five, six, seven, eight, nine, ten, eleven or twelve different antigenic peptides or proteins originating from the same RSV and / or a different RSV.

[0289] In one embodiment, the composition may comprise at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or even more different artificial mRNAs, as defined in connection with the first aspect of the invention, each encoding at least one antigenic peptide or protein derived from genetically the same RSV or a fragment or variant thereof. The terms "the same," "identical," or "same RSV," used in connection with a virus, e.g., "the same virus," are to be understood as genetically identical. In particular, the (genetically) identical virus expresses the same proteins or peptides, wherein all proteins or peptides have the same amino acid sequence. Specifically, the (genetically) identical RSV expresses substantially the same proteins, peptides, or polyproteins, wherein these proteins, peptides, or polyproteins preferably do not differ in their amino acid sequence(s).A non-restrictive list of exemplary RSV viruses is provided in List 1.

[0290] In embodiments, the composition comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more different mRNAs, as defined in connection with the first aspect of the invention, each encoding at least one peptide or protein derived from a genetically distinct RSV or a fragment or variant thereof. The terms "different," "different," or "other RSV," as used in this description in connection with a virus, e.g., an "other" virus, are to be understood as the difference between at least two corresponding viruses, the difference being manifested on the RNA genome of the respective other virus. In particular, the (genetically) different RSV expresses at least one different protein, peptide, or polyprotein, the at least one different protein, peptide, or polyprotein preferably differing in at least one amino acid.

[0291] In other embodiments, the composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more additional mRNA constructs encoding RSV antigens selected from glycoprotein G, short hydrophobic protein SH, matrix protein M, nucleoprotein N, large polymerase L, M2-1 protein, M2-2 protein, phosphoprotein P, non-structural protein NS1 or non-structural protein NS2 or any combination thereof.

[0292] To produce a composition comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 further mRNA constructs encoding RSV, methods are preferably used as disclosed in PCT application PCT / EP2016 / 082487 or in published patent application WO2017 / 1090134A1, and adapted accordingly.

[0293] In preferred embodiments, the composition of the second aspect comprises at least one artificial mRNA of the first aspect and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, selected from matrix protein M, nucleoprotein N, M2-1 protein and / or phosphoprotein P or combinations thereof. The addition of a further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, N, M2-1 and / or phosphoprotein P (or combinations thereof) is particularly advantageous, for example, to promote a T-cell immune response.

[0294] In particular, embodiments relating to the artificial mRNA of the first aspect can also be read and understood as suitable embodiments of the at least one further artificial RNA of the second aspect (e.g. embodiments relating to UTR combinations, cds optimizations, histone stem loop, PolyA, PolyC, cap structure, mRNA structure, mRNA production and mRNA purification, etc.).

[0295] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from the RSV matrix protein M.

[0296] Alternatively, the composition of the second aspect may conveniently include at least one artificial mRNA of the first aspect encoding RSV F and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0297] Alternatively, the composition of the second aspect may suitably include at least one artificial mRNA of the first aspect encoding RSV F and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0298] Alternatively, the composition of the second aspect may appropriately include at least one artificial mRNA of the first aspect encoding RSV F and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0299] In preferred embodiments, the composition of the second aspect comprises two further artificial RNA species, each comprising at least one coding sequence encoding at least one RSV-derived antigenic peptide or protein selected from matrix protein M, nucleoprotein N, M2-1 and phosphoprotein P.

[0300] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0301] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0302] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0303] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0304] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0305] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0306] In preferred embodiments, the composition of the second aspect comprises three further artificial RNA species, each comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV and selected from matrix protein M, nucleoprotein N, M2-1 and phosphoprotein P.

[0307] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0308] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0309] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0310] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0311] Accordingly, the composition of the second aspect may expediently include at least one artificial mRNA of the first aspect encoding RSV F and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0312] In preferred embodiments, the composition of the second aspect comprises four further artificial RNA species, each comprising at least three further artificial RNA species, each comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV and selected from matrix protein M, nucleoprotein N, M2-1 and phosphoprotein P.

[0313] Accordingly, the composition of the second aspect may expediently include at least one artificial RNA of the first aspect encoding RSV F, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein originating from RSV M, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein originating from RSV N, at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein originating from RSV P, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein originating from RSV M2-1.

[0314] In preferred embodiments of the second aspect, the coding sequence of the further artificial RNA encodes at least one of the amino acid sequences that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to one of the SEQ ID NOs: 9684, 10053-10133, 10134, 10503-10636, 10637, 11006-11182, 11183, 11552-11725, 19844, 20213, 20582, 20951 or a fragment or variant of one of these Sequences. Additional information on each of these suitable amino acid sequences encoding RSV proteins can also be obtained from the sequence protocol, in particular from the entries therein under the identifier <223> The details are as follows.

[0315] In further embodiments of the second aspect, the coding sequence of the additional artificial RNA encodes at least one antigenic peptide or protein, as defined herein, and additionally a heterologous secretory signal sequence or a heterologous secretory signal peptide. The heterologous secretory signal sequence can increase the secretion of the encoded antigenic peptide or protein.

[0316] Suitable secretory signal peptides can be selected from the list of amino acid sequences according to SEQ ID NOs: 1-1115 and SEQ ID NO: 1728 of patent application WO2017 / 081082, or from fragments or variants of these sequences. At the nucleic acid level, any nucleic acid sequence (e.g., RNA sequence) that encodes such amino acid sequences can be selected. In this context, the disclosure of WO2017 / 081082 is hereby incorporated by reference.

[0317] According to embodiments, the secretory signal sequence comprises an amino acid sequence that is identical to, or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to, one of the SEQ ID NOs: 21329-21362 or a fragment or variant of one of these sequences. Additional information on each of these suitable amino acid sequences encoding secretory signal sequences can also be found in the sequence protocol, in particular the entries therein under the identifier <223> details provided.

[0318] In preferred embodiments, the further artificial RNA of the composition comprises a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV M, N, M2-1 and / or phosphoprotein P or a fragment or variant thereof, and preferably encoding an amino acid sequence selected from one of the SEQ ID NOs: 9684, 10053-10133, 10134, 10503-10636; 10637, 11006-11182, 11183, 11552-11725, 19844, 20213, 20582, 20951 or a fragment or variant of one of these sequences.

[0319] In preferred embodiments, the further artificial RNA of the composition comprises a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV M2-1 or a fragment or variant thereof, preferably encoding an amino acid sequence selected from one of the SEQ ID NOs: 11183, 20953, 21414 or a fragment or variant of one of these sequences.

[0320] Advantageously, the further artificial RNA of the composition comprises a coding sequence functionally linked to a 3'-UTR and a 5'-UTR selected from a-1, a-2, a-3, a-4, a-5, b-1, b-2, b-3, b-4, b-5, c-1, c-2, c-3, c-4, c-5, d-1, d-2, d-3, d-4, d-5, e-1, e-2, e-3, e-4, e-5, e-6, f-1, f-2, f-3, f-4, f-5, g-1, g-2, g-3, g-4, g-5, h-1, h-2, h-3, h-4, h-5, i-1, i-2, or i-3 (as defined in connection with the first aspect).

[0321] Advantageously, the further artificial RNA of the composition comprises a coding sequence in which at least one or more than one or preferably all uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0322] Accordingly, in other embodiments, the further artificial RNA of the composition may comprise a 5'-cap sequence element according to SEQ ID NOs: 43 or 21321, or a fragment or variant thereof.

[0323] In preferred embodiments of the second aspect, the coding sequence of the further RNA comprises at least one of the nucleic acid sequences that are identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11119, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21385-21388, 21411-21414 or a fragment or variant of any of these sequences (cds optimizations as defined in connection with the first aspect). Additional information on each of these suitable amino acid sequences encoding RSV proteins can also be found in the sequence protocol, in particular under the identifier <223> The details are as follows.

[0324] In preferred embodiments of the second aspect, the further artificial RNA, preferably the further mRNA of the composition, preferably in the 5' to 3' direction, comprises the following elements a) -i): a) 5'-cap structure, preferably as indicated in connection with the first aspect, most preferably a Cap1 structure; b) optionally a 5'-UTR as specified in connection with the first aspect, preferably at least one selected from SEQ ID NOs: 1-22; c) a ribosome binding site, preferably as specified here; d) at least one coding sequence selected from SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11119, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21385-21388, 21411-21414; e) 3'-UTR as specified in connection with the first aspect, preferably at least one selected from SEQ ID NOs: 23-38; f) optionally a poly(A) sequence, preferably as indicated in connection with the first aspect; g) optionally a poly(C) sequence, preferably as specified in connection with the first aspect; h) optionally a histone stem loop, preferably as specified in connection with the first aspect; i) optionally a 3'-terminal sequence element as specified in connection with the first aspect, preferably according to SEQ ID NOs: 44-63, or 21322-21328, and wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0325] In particularly preferred embodiments of the second aspect, the further artificial RNA, preferably the further mRNA of the composition, preferably in the 5' to 3' direction comprises the following elements a) -i): a) 5'-cap structure, preferably as indicated in connection with the first aspect, most preferably a Cap1 structure; b) optionally 5'-UTR as indicated in connection with the first aspect, preferably at least one selected from SEQ ID NOs: 1-22; c) a ribosome binding site, preferably as specified here; d) at least one coding sequence selected from SEQ ID NOs: 11185-11191, 21388, 20952-20959, 21414; e) 3'-UTR as specified in connection with the first aspect, preferably at least one selected from SEQ ID NOs: 23-38; f) optionally a poly(A) sequence, preferably as indicated in connection with the first aspect; g) optionally a poly(C) sequence, preferably as specified in connection with the first aspect; h) optionally a histone stem loop, preferably as specified in connection with the first aspect; i) optionally a 3'-terminal sequence element as specified in connection with the first aspect, preferably according to SEQ ID NOs: 44-63, or 21322-21328, and wherein optionally at least one or more than one, preferably all, uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0326] In this context, the preferred RSV polypeptide, nucleic acid and mRNA sequences of the second aspect are listed in Table 7A.

[0327] In Table 7A, columns A to D represent specific suitable constructs of the second aspect derived from the RSV protein, with column A providing suitable sequences for M, column B suitable sequences for N, column C suitable sequences for P and column D suitable sequences for M2-1.

[0328] The protein name designs are given in line 1 (columns A to D), and the protein SEQ ID numbers given in the sequence protocol are given in line 2 (“PRT”). Preferred SEQ ID NOs of HRSV(A2) are given in line 3 (“HRSV(A2) PRT”). The SEQ ID NOs of the corresponding coding sequences for each specified HRSV(A2) protein are given in lines 4-11 (“cds wt”, “cds opt1”, “cds opt2”, “cds opt3”, “cds opt4”, “cds opt5”, “cds opt6”, “cds opt11”). The SEQ ID NOs of the corresponding mRNA constructs comprising the coding sequences and suitable 3' and 5' UTRs according to the invention are given in lines 12 to 55 (“mRNA design a-1” to “mRNA design i-3”, as shown herein).

[0329] Preferred protein SEQ ID NOs of HRSV(Memphis-37) are given in line 56 (“Memphis-37 PRT”). The SEQ ID NOs of the corresponding coding sequences for each specified HRSV(Memphis-37) protein are given in lines 57-64 (“cds wt”, “cds opt1”, “cds opt2”, “cds opt3”, “cds opt4”, “cds opt5”, “cds opt6”, and “cds opt11”, respectively). The SEQ ID NOs of the corresponding mRNA constructs comprising the coding sequences and suitable 3' and 5' UTRs according to the invention are given in lines 65 to 108 (“mRNA design a-1” to “mRNA design i-3”, as specified herein).

[0330] Further information, e.g., about the type of coding sequence contained in the specified mRNA constructs, can be found in the <223> -Identifier of the respective SEQ ID NO included in the sequence protocol. Table 7A: Preferred additional coding sequences and mRNA constructs of the composition or vaccine 1 Stamm Beschreibung A B C D RSV M RSV N RSV P RSV M2-1 2 Divers PRT 10053-10133 10503-10636 11006-11182 11552-11725 3 HRSV(A2) PRT 9684 10134 10637 11183 4 HRSV(A2) KodierendeSequenz (cds) WildTyp 9685 10135 10638 11184 5 HRSV(A2) CDS opt1 9686, 21385 10136, 21386 10639, 21387 11185, 21388 6 HRSV(A2) cds opt2 9687 10137 10640 11186 7 HRSV(A2) cds opt3 9688 10138 10641 11187 8 HRSV(A2) cds opt4 9689 10139 10642 11188 9 HRSV(A2) cds opt5 9690 10140 10643 11189 10 HRSV(A2) cds opt6 9691 10141 10644 11190 11 HRSV(A2) cds opt11 9692 10142 10645 11191 12 HRSV(A2) mRNA Design a-1 9693-9700,21481, 21482,21627, 21628 10143-10150,21483, 21484,21629, 21630 10646-10653,21485, 21486,21631, 21632 11192-11199,21487, 21488,21633, 21634 13 HRSV(A2) mRNA Design a-2 9701-9708 10151-10158 10654-10661 11200-11207 14 HRSV(A2) mRNA Design a-3 9709-9716 10159-10166 10662-10669 11208-11215 15 HRSV(A2) mRNA Design a-4 9717-9724 10167-10174 10670-10677 11216-11223 16 HRSV(A2) mRNA Design a-5 9725-9732 10175-10182 10678-10685 11224-11231 17 HRSV(A2) mRNA Design b-1 9733-9740 10183-10190 10686-10693 11232-11239 18 HRSV(A2) mRNA Design b-2 9741-9748 10191-10198 10694-10701 11240-11247 19 HRSV(A2) mRNA Design b-3 9749-9756 10199-10206 10702-10709 11248-11255 20 HRSV(A2) mRNA Design b-4 9757-9764 10207-10214 10710-10717 11256-11263 21 HRSV(A2) mRNA Design b-5 9765-9772 10215-10222 10718-10725 11264-11271 22 HRSV(A2) mRNA Design c-1 9773-9780 10223-10230 10726-10733 11272-11279 23 HRSV(A2) mRNA Design c-2 9781-9788 10231-10238 10734-10741 11280-11287 24 HRSV(A2) mRNA Design c-3 9789-9796 10239-10246 10742-10749 11288-11295 25 HRSV(A2) mRNA Design c-4 9797-9804 10247-10254 10750-10757 11296-11303 26 HRSV(A2) mRNA Design c-5 9805-9812 10255-10262 10758-10765 11304-11311 27 HRSV(A2) mRNA Design d-1 9813-9820 10263-10270 10766-10773 11312-11319 28 HRSV(A2) mRNA Design d-2 9821-9828 10271-10278 10774-10781 11320-11327 29 HRSV(A2) mRNA Design d-3 9829-9836 10279-10286 10782-10789 11328-11335 30 HRSV(A2) mRNA Design d-4 9837-9844 10287-10294 10790-10797 11336-11343 31 HRSV(A2) mRNA Design d-5 9845-9852 10295-10302 10798-10805 11344-11351 32 HRSV(A2) mRNA Design e-1 9853-9860 10303-10310 10806-10813 11352-11359 33 HRSV(A2) mRNA Design e-2 9861-9868 10311-10318 10814-10821 11360-11367 34 HRSV(A2) mRNA Design e-3 9869-9876 10319-10326 10822-10829 11368-11375 35 HRSV(A2) mRNA Design e-4 9877-9884 10327-10334 10830-10837 11376-11383 36 HRSV(A2) mRNA Design e-5 9885-9892 10335-10342 10838-10845 11384-11391 37 HRSV(A2) mRNA Design e-6 9893-9900 10343-10350 10846-10853 11392-11399 38 HRSV(A2) mRNA Design f-1 9901-9908 10351-10358 10854-10861 11400-11407 39 HRSV(A2) mRNA Design f-2 9909-9916 10359-10366 10862-10869 11408-11415 40 HRSV(A2) mRNA Design f-3 9917-9924 10367-10374 10870-10877 11416-11423 41 HRSV(A2) mRNA Design f-4 9925-9932 10375-10382 10878-10885 11424-11431 42 HRSV(A2) mRNA Design f-5 9933-9940 10383-10390 10886-10893 11432-11439 43 HRSV(A2) mRNA Design g-1 9941-9948 10391-10398 10894-10901 11440-11447 44 HRSV(A2) mRNA Design g-2 9949-9956 10399-10406 10902-10909 11448-11455 45 HRSV(A2) mRNA Design g-3 9957-9964 10407-10414 10910-10917 11456-11463 46 HRSV(A2) mRNA Design g-4 9965-9972 10415-10422 10918-10925 11464-11471 47 HRSV(A2) mRNA Design g-5 9973-9980 10423-10430 10926-10933 11472-11479 48 HRSV(A2) mRNA Design h-1 9981-9988 10431-10438 10934-10941 11480-11487 49 HRSV(A2) mRNA Design h-2 9989-9996 10439-10446 10942-10949 11488-11495 50 HRSV(A2) mRNA Design h-3 9997-10004 10447-10454 10950-10957 11496-11503 51 HRSV(A2) mRNA Design h-4 10005-10012 10455-10462 10958-10965 11504-11511 52 HRSV(A2) mRNA Design h-5 10013-10020 10463-10470 10966-10973 11512-11519 53 HRSV(A2) mRNA Design i-1 10021-10028 10471-10478 10974-10981 11520-11527 54 HRSV(A2) mRNA Design i-2 10029-10044 10479-10494 10982-10997 11528-11543 55 HRSV(A2) mRNA Design i-3 10045-10052 10495-10502 10998-11005 11544-11551 56 Memphis-37 PRT 19844 20213 20582 20951 57 Memphis-37 CDS wt 19845 20214 20583 20952 58 Memphis-37 CDS opt1 19846, 21411 20215, 21412 20584, 21413 20953, 21414 59 Memphis-37 CDS opt2 19847 20216 20585 20954 60 Memphis-37 CDS opt3 19848 20217 20586 20955 61 Memphis-37 CDS opt4 19849 20218 20587 20956 62 Memphis-37 CDS opt5 19850 20219 20588 20957 63 Memphis-37 CDS opt6 19851 20220 20589 20958 64 Memphis-37 CDS opt11 19852 20221 20590 20959 65 Memphis-37 mRNA Design a-1 19853-19860, 20222-20229, 20591-20598, 20960-20967, 21553, 21554, 21555, 21556, 21557, 21558, 21559, 21560, 21699, 21700 21701, 21702 21703, 21704 21705, 21706 66 Memphis-37 mRNA Design a-2 19861-19868 20230-20237 20599-20606 20968-20975 67 Memphis-37 mRNA Design a-3 19869-19876 20238-20245 20607-20614 20976-20983 68 Memphis-37 mRNA Design a-4 19877-19884 20246-20253 20615-20622 20984-20991 69 Memphis-37 mRNA Design a-5 19885-19892 20254-20261 20623-20630 20992-20999 70 Memphis-37 mRNA Design b-1 19893-19900 20262-20269 20631-20638 21000-21007 71 Memphis-37 mRNA Design b-2 19901-19908 20270-20277 20639-20646 21008-21015 72 Memphis-37 mRNA Design b-3 19909-19916 20278-20285 20647-20654 21016-21023 73 Memphis-37 mRNA Design b-4 19917-19924 20286-20293 20655-20662 21024-21031 74 Memphis-37 mRNA Design b-5 19925-19932 20294-20301 20663-20670 21032-21039 75 Memphis-37 mRNA Design c-1 19933-19940 20302-20309 20671-20678 21040-21047 76 Memphis-37 mRNA Design c-2 19941-19948 20310-20317 20679-20686 21048-21055 77 Memphis-37 mRNA Design c-3 19949-19956 20318-20325 20687-20694 21056-21063 78 Memphis-37 mRNA Design c-4 19957-19964 20326-20333 20695-20702 21064-21071 79 Memphis-37 mRNA Design c-5 19965-19972 20334-20341 20703-20710 21072-21079 80 Memphis-37 mRNA Design d-1 19973-19980 20342-20349 20711-20718 21080-21087 81 Memphis-37 mRNA Design d-2 19981-19988 20350-20357 20719-20726 21088-21095 82 Memphis-37 mRNA Design d-3 19989-19996 20358-20365 20727-20734 21096-21103 83 Memphis-37 mRNA Design d-4 19997-20004 20366-20373 20735-20742 21104-21111 84 Memphis-37 mRNA Design d-5 20005-20012 20374-20381 20743-20750 21112-21119 85 Memphis-37 mRNA Design e-1 20013-20020 20382-20389 20751-20758 21120-21127 86 Memphis-37 mRNA Design e-2 20021-20028 20390-20397 20759-20766 21128-21135 87 Memphis-37 mRNA Design e-3 20029-20036 20398-20405 20767-20774 21136-21143 88 Memphis-37 mRNA Design e-4 20037-20044 20406-20413 20775-20782 21144-21151 89 Memphis-37 mRNA Design e-5 20045-20052 20414-20421 20783-20790 21152-21159 90 Memphis-37 mRNA Design e-6 20053-20060 20422-20429 20791-20798 21160-21167 91 Memphis-37 mRNA Design f-1 20061-20068 20430-20437 20799-20806 21168-21175 92 Memphis-37 mRNA Design f-2 20069-20076 20438-20445 20807-20814 21176-21183 93 Memphis-37 mRNA Design f-3 20077-20084 20446-20453 20815-20822 21184-21191 94 Memphis-37 mRNA Design f-4 20085-20092 20454-20461 20823-20830 21192-21199 95 Memphis-37 mRNA Design f-5 20093-20100 20462-20469 20831-20838 21200-21207 96 Memphis-37 mRNA Design g-1 20101-20108 20470-20477 20839-20846 21208-21215 97 Memphis-37 mRNA Design g-2 20109-20116 20478-20485 20847-20854 21216-21223 98 Memphis-37 mRNA Design g-3 20117-20124 20486-20493 20855-20862 21224-21231 99 Memphis-37 mRNA Design g-4 20125-20132 20494-20501 20863-20870 21232-21239 100 Memphis-37 mRNA Design g-5 20133-20140 20502-20509 20871-20878 21240-21247 101 Memphis-37 mRNA Design h-1 20141-20148 20510-20517 20879-20886 21248-21255 102 Memphis-37 mRNA Design h-2 20149-20156 20518-20525 20887-20894 21256-21263 103 Memphis-37 mRNA Design h-3 20157-20164 20526-20533 20895-20902 21264-21271 104 Memphis-37 mRNA Design h-4 20165-20172 20534-20541 20903-20910 21272-21279 105 Memphis-37 mRNA Design h-5 20173-20180 20542-20549 20911-20918 21280-21287 106 Memphis-37 mRNA Design i-1 20181-20188 20550-20557 20919-20926 21288-21295 107 Memphis-37 mRNA Design i-2 20189-20204 20558-20573 20927-20942 21296--21311 108 Memphis-37 mRNA Design i-3 20205-20212 20574-20581 20943-20950 21312-21319

[0331] Accordingly, in preferred embodiments, the composition of the second aspect comprises at least one further artificial RNA comprising at least one coding sequence encoding at least one RSV-derived antigenic peptide or protein selected from matrix protein M, nucleoprotein N, M2-1 protein, phosphoprotein P, wherein the further artificial RNA comprises or consists of an RNA sequence that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence composed of SEQ ID NOs: 9693-10052, 10143-10502, 10646-11005, 11192-11551, 19853-20212, 20222-20581, 20591-20950, 20960-21319, 21481-21488, 21627-21634, 21553-21560, 21699-21706 or a fragment or variant of one of these sequences is selected, wherein, where appropriate,at least one or more than one or alfe uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides. Additional information on each of these suitable amino acid sequences encoding RSV proteins can also be found in the sequence protocol, in particular in the entries there under the identifier <223> The information provided is explained below.

[0332] In particularly preferred embodiments of the second aspect, the composition of the second aspect comprises - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and another artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, 21700 (encoded RSV M); or - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and another artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10143-10502, 20222-20581, 21483, 21484, 21629, 21630, 21555, 21556, 21701, 21702 (encoded RSV N); or - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and at least one other artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, 21704 (coding for RSV P); or - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and at least one other artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11192-11551, 20960-21319, 21487, 21488, 21633, 21634, 21559, 21560, 21705, 21706 (RSV M2-1 encoding); or - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and at least one other artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, 21700 (encoding RSV M) and another artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, 21704 (encoding RSV P); - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and at least one other artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, 21700 (encoding RSV M) and another artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, 21704 (encoding RSV P) and another artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 10143-10502, 20222-20581, 21483, 21484, 21629, 21630, 21555, 21556, 21701, 21702 (coding for RSV N), or, - at least one artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21526-21528, 21582-21584, 21588-21590, 21594-21596, 21600-21602, 21672-21674 (encoding RSV F, as defined in the first aspect) and at least one other artificial RNA that is identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11192-11551, 20960-21319, 21487, 21488, 21633, 21634, 21559, 21560, 21705, 21706 (RSV M2-1 encoding), where, if applicable,at least one or more than one or all uracil nucleotides are replaced by pseudouridine (ψ) nucleotides or N1-methylpseudouridine (m1ψ) nucleotides.

[0333] In various embodiments, different combinations of RSV F (F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4) RNA constructs and RSV T-cell antigen RNA constructs (RSV M, N, M2-1 or P) can suitably be included in the composition (as disclosed in Table 7B; combinations 1-64). Combinations 49-64 are preferred, with combination 64 being particularly preferred. Table 7B: Claimed and other combinations of RNA constructs encoding RSV F and RNA constructs encoding T-cell antigens Kombination RSV F Konstrukt SEQ ID NO:Protein T-ZellantigenKonstrukt SEQ ID NO:Protein 1 F0 68 M 9684 2 F-del 483 M 9684 3 F_DSCav1 898 M 9684 4 F-del_DSCav1 1267 M 9684 5 F_DSCav1_mut1 1636 M 9684 6 F-del_DSCav1_mut1 2005 M 9684 7 F_DSCav1_mut2 2374 M 9684 8 F-del_DSCav1_mut2 2743 M 9684 9 F_DSCav1_mut3 3112 M 9684 10 F-del_DSCav1_mut3 3481 M 9684 11 F_DSCav1_mut0 3850 M 9684 12 F-del_DSCav1_mut0 4219 M 9684 13 F_DSCav1_mut4 4588 M 9684 14 F-del_DSCav1_mut4 4957 M 9684 15 F_DSCav1_mut5 5326 M 9684 16 F-del_DSCav1_mut5 5695 M 9684 17 F0 68 N 10134 18 F-del 483 N 10134 19 F_DSCav1 898 N 10134 20 F-del_DSCav1 1267 N 10134 21 F_DSCav1_mut1 1636 N 10134 22 F-del_DSCav1_mut1 2005 N 10134 23 F_DSCav1_mut2 2374 N 10134 24 F-del_DSCav1_mut2 2743 N 10134 25 F_DSCav1_mut3 3112 N 10134 26 F-del_DSCav1 _ mut3 3481 N 10134 27 F_DSCav1_mut0 3850 N 10134 28 F-del_DSCav1_mut0 4219 N 10134 29 F_DSCav1_mut4 4588 N 10134 30 F-del_DSCav1_mut4 4957 N 10134 31 F_DSCav1_mut5 5326 N 10134 32 F-del_DSCav1_mut5 5695 N 10134 33 F0 68 P 10637 34 F-del 483 P 10637 35 F_DSCav1 898 P 10637 36 F-del_DSCav1 1267 P 10637 37 F_DSCav1_mut1 1636 P 10637 38 F-del_DSCav1_mut1 2005 P 10637 39 F_DSCav1_to2 2374 P 10637 40 F-part_DSCav1_mut2 2743 P 10637 41 F_DSCav1_to3 3112 P 10637 42 F-part_DSCav1_mut3 3481 P 10637 43 F_DSCav1_to0 3850 P 10637 44 F-part_DSCav1_mut0 4219 P 10637 45 F_DSCav1_to4 4588 P 10637 46 Part F_DSCav1_mut4 4957 P 10637 47 F_DSCav1_to5 5326 P 10637 48 Part F_DSCav1_mut5 5695 P 10637 49 F0 68 M2-1 11183 50 Part F 483 M2-1 11183 51 F_DSCav1 898 M2-1 11183 52 Part F_DSCav1 1267 M2-1 11183 53 F_DSCav1_to1 1636 M2-1 11183 54 F-part_DSCav1_mut1 2005 M2-1 11183 55 F_DSCav1_to2 2374 M2-1 11183 56 Part F DSCav1 mut2 2743 M2-1 11183 57 F_DSCav1_to3 3112 M2-1 11183 58 F-part_DSCav1_mut3 3481 M2-1 11183 59 F_DSCav1_to0 3850 M2-1 11183 60 F-part_DSCav1_mut0 4219 M2-1 11183 61 F_DSCav1_to4 4588 M2-1 11183 62 Part F_DSCav1_mut4 4957 M2-1 11183 63 F_DSCav1_to5 5326 M2-1 11183 64 Part F_DSCav1_mut5 5695 M2-1 11183

[0334] Table 7B discloses compositions of the second aspect, comprising at least one RNA encoding RSV F from strain A2, as defined in the first aspect, and at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV of strain A2, selected from matrix protein M, nucleoprotein N, M2-1 protein, and phosphoprotein P. In further embodiments of the invention, all compositions disclosed in Table 7B are also applicable to antigenic peptides or proteins derived from an RSV isolate Memphis-37 (strain Memphis-37).

[0335] In a particularly preferred embodiment, the composition comprises an RSV F RNA construct (F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4) and additionally an RSV M2-1 RNA construct, preferably F-del_DSCav1_mut5 and M2-1.

[0336] In various embodiments, the at least one artificial RNA of the first aspect and the at least one further artificial RNA, as specified here, are derived from the same RSV virus (e.g., any virus from List 1).

[0337] In preferred embodiments, the at least one artificial RNA of the first aspect is derived from HRSV(A2) and the at least one further artificial RNA, as specified here, is derived from HRSV(A2).

[0338] In preferred embodiments, the at least one artificial RNA of the first aspect is derived from HRSV(Memphis-37) and the at least one further artificial RNA, as specified here, is derived from HRSV(Memphis-37).

[0339] It is understood that, within the scope of the invention, certain combinations of coding sequences can be generated by any combination of monocistronic, bicistronic and multicistronic artificial nucleic acids and / or multi-antigen constructs / nucleic acids to obtain a nucleic acid composition that codes for several antigenic peptides or proteins, as defined herein.

[0340] Furthermore, one or more compatible solid or liquid excipients, diluents, or encapsulating compounds suitable for administration to humans may be used. The term "compatible" as used here means that the components of the composition can be mixed with the at least one RNA and, where applicable, the other artificial RNA of the composition in such a way that no interactions occur that would substantially reduce the biological activity or pharmaceutical efficacy of the composition under typical conditions of use. Pharmaceutically acceptable carriers, excipients, and diluents must have sufficiently high purity and sufficiently low toxicity to be suitable for administration to a patient.Compounds that can be used as pharmaceutically acceptable carriers, fillers, or components thereof include sugars, such as lactose, glucose, trehalose, and sucrose; starches, such as corn starch or potato starch; dextrose; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; tallow; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and theobroma oil; polyols, such as polypropylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; and alginic acid.

[0341] Other additives that may be included in the composition are emulsifiers, such as tween, wetting agents, such as sodium lauryl sulfate, colorants, flavor enhancers, pharmaceutical carriers, tablet formers, stabilizers, antioxidants and preservatives. Complexity:

[0342] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated, or at least partially complexed or partially associated, with one or more cationic or polycationic compounds, preferably a cationic or polycationic polymer, a cationic or polycationic polysaccharide, a cationic or polycationic lipid, a cationic or polycationic protein, a cationic or polycationic peptide or any combination thereof.

[0343] The term “cationic or polycationic compound,” as used here, is recognized and understood by those skilled in the art and refers, for example, to a charged molecule that is positively charged at a pH in the range of about 1 to 9, at a pH in the range of about 3 to 8, at a pH in the range of about 4 to 8, at a pH in the range of about 5 to 8, preferably at a pH in the range of about 6 to 8, more preferably at a pH in the range of about 7 to 8, and most preferably at a physiological pH, e.g., in the range of about 7.2 to about 7.5. Accordingly, a cationic component, e.g., a cationic peptide, a cationic protein, a cationic polymer, a cationic polysaccharide, or a cationic lipid, can be any positively charged compound or polymer that is positively charged under physiological conditions.A “cationic or polycationic peptide or protein” can contain at least one positively charged amino acid or more than one positively charged amino acid, e.g., selected from Arg, His, Lys, or Orn. ​​Accordingly, “polycationic” components that exhibit more than one positive charge under the given conditions also fall within the scope of application.

[0344] Cationic or polycationic compounds that are particularly preferred in this context may be selected from the following list of cationic or polycationic peptides or proteins or fragments thereof: protamine, nucleolin, spermine or spermidine, or other cationic peptides or proteins, such as poly-L-lysine (PLL), poly-arginine, basic polypeptides, cell-penetrating peptides (CPPs), including HIV-binding peptides, HIV-1 Tat (HIV), Tat-derived peptides, penetratin, VP22-derived or analogous peptides, HSV VP22 (herpes simplex), MAP, KALA or protein transduction domains (PTDs), PpT620, proline-rich peptides, arginine-rich peptides, lysine-rich peptides, MPG peptide(s), Pep-1, L-oligomers, calcitonin peptide(s). Antennapedia-derived peptides, pAntp, plsl, FGF, Lactoferrin, Transportan, Buforin-2, Bac715-24, SynB, SynB(1), pVEC, hCT-derived peptides, SAP, or histones.Even more preferably, the nucleic acid defined herein, preferably the mRNA defined herein, is complexed with one or more polycations, preferably with protamine or oligofectamine, particularly preferably with protamine.

[0345] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and, optionally, the further artificial RNA of the second aspect is complexed with protamine.

[0346] Other preferred cationic or polycationic compounds that can be used as transfection or complexing agents may include cationic polysaccharides, e.g., chitosan, polybrene, etc.; cationic lipids, e.g., DOTMA, DMRIE, Di-C14-amidin, DOTIM, SAINT, DC-Chol, BGTC, CTAP, DOPC, DODAP, DOPE: Dioleyl-Phosphatidylethanol-Amine, DOSPA, DODAB, DOIC, DMEPC, DOGS, DIMRI, DOTAP, DC-6-14, CLIP1, CLIP6, CLIP9, Oligofectamin; or cationic or polycationic polymers, e.g., modified polyamino acids such as beta-amino acid polymers or reverse polyamides, etc., modified polyethylenes such as PVP, etc., modified acrylates such as pDMAEMA, etc., modified amidoamines such as pAMAM, etc., modified polybetaamino esters (PBAE) such as diamine-terminal modified 1,4-butanediol diacrylate-co-5-amino-1-pentanol polymers, etc., dendrimers such as polypropylamine dendrimers or pAMAM-based dendrimers, etc., polyimines such as PEI, poly(propylenimine), etc., Polyallylamine, sugar-based polymers such as cyclodextrin-based polymers, dextran-based polymers, etc., silane-backbone-based polymers such as PMOXA-PDMS copolymers, etc., block polymers consisting of a combination of one or more cationic blocks (e.g., selected from a cationic polymer as above) and one or more hydrophilic or hydrophobic blocks (e.g., polyethylene glycol); etc.

[0347] In this context, it is particularly preferred that the at least one artificial RNA, as defined herein, and optionally the further artificial RNA of the second aspect, is complexed or at least partially complexed with a cationic or polycationic compound and / or a polymeric support, preferably cationic proteins or peptides. Partially means that only a portion of the artificial nucleic acid is complexed with a cationic compound and that the remainder of the artificial nucleic acid (contained in the (pharmaceutical) composition according to the invention) is in uncomplexed form ("free").

[0348] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed with one or more cationic or polycationic compounds, preferably protamine, and at least one free artificial RNA of the first aspect and optionally the further RNA of the second aspect.

[0349] In this context, it is particularly preferred that the at least one artificial RNA, as defined herein, and optionally the further artificial RNA of the second aspect, is complexed or at least partially complexed with protamine. Preferably, the molar ratio of the nucleic acid, in particular the RNA of the protamine-complexed RNA, to the free RNA can be selected from a molar ratio of about 0.001:1 to about 1:0.001, including a ratio of about 1:1. Advantageously, the complexed RNA is complexed with protamine by adding a protamine-trehalose solution to the RNA sample in a weight (w / w) RNA:protamine ratio of 2:1.

[0350] Further preferred cationic or polycationic proteins or peptides that can be used for complexation can be derived from the formula (Arg)I;(Lys)m;(His)n;(Orn)o;(Xaa)x of patent application WO2009 / 030481 or WO2011 / 026641.

[0351] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or at least partially complexed with at least one cationic or polycationic protein or peptide, preferably selected from SEQ ID NOs: 64-67, 21320 or any combination thereof.

[0352] According to embodiments, the composition of the present invention comprises the RNA defined herein and a polymeric carrier.

[0353] The term "polymeric carrier," as used here, is recognized and understood by those skilled in the art and refers, for example, to a compound that facilitates the transport and / or complexation of another compound (cargo). A polymeric carrier is typically a carrier formed from a polymer. A polymeric carrier can be bonded to its charge (nucleic acid, RNA) through covalent or non-covalent interactions.

[0354] A suitable polymeric support can be a polymeric support consisting of disulfide-crosslinked cationic compounds. The disulfide-crosslinked cationic compounds can be identical or different from one another. The polymeric support can also contain further components. The polymeric support used according to the invention can contain mixtures of cationic peptides, proteins, or polymers and optionally further components, as defined herein, which are crosslinked by disulfide bonds (via -SH groups).

[0355] In this context, polymeric carriers according to the formula {(Arg)I;(Lys)m;(His)n;(Orn)o;(Xaa')x(Cys)y} and the formula Cys,{(Arg)I;(Lys)m;(His)n;(Om)o;(Xaa)x}Cys2 of patent application WO2012 / 013326 are preferred.

[0356] In embodiments, the polymeric support used to complex the RNA defined herein can be a polymeric support molecule according to the formula (LP 1 -S-[SP2 -S] n -SP 3 -L) derived from patent application WO2011 / 026641.

[0357] In embodiments, the polymeric support compound is formed by, comprised of, or consists of the peptide elements CysArg12Cys (SEQ ID NO: 64), CysArg12 (SEQ ID NO: 65), or TrpArg12Cys (SEQ ID NO: 66). In particularly preferred embodiments, the polymeric support compound consists of a (R 12 C)-(R 12 C)-dimer, a (WR 12 C)-(WR 12 C)-dimer or a (CR 12 )-(CR 12 C)-(CR 12 )-Trimer, wherein the individual peptide elements in the dimer (e.g. (WR12C)) or the trimer (e.g. (CR12)) are linked via -SH groups.

[0358] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CHHHHHRRRRRHHHHHHC-S-)7-S-PEG5000-OH (SEQ ID NO: 67 as peptide monomer).

[0359] In a further preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CHHHHHRRRRRHHHHHHC-S-)4-S-PEG5000-OH (SEQ ID NO: 67 as peptide monomer).

[0360] In a further preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CGHHHHHRRRRHHHHHGC-S-)7-S-PEG5000-OH (SEQ ID NO: 21320 as peptide monomer).

[0361] In a further preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CGHHHHHRRRRHHHHHGC-S-)4-S-PEG5000-OH (SEQ ID NO: 21320 as peptide monomer).

[0362] In other embodiments, the composition comprises at least one artificial RNA as described herein and optionally the further artificial RNA of the second aspect, wherein the at least one artificial RNA and optionally the further artificial RNA of the second aspect is complexed or associated with polymeric supports and optionally with at least one lipid component, as described in the published PCT applications WO2017 / 212008A1, WO2017 / 212006A1, WO2017 / 212007A1 and WO2017 / 212009A1.

[0363] In a particularly preferred embodiment, the polymeric support is a peptide polymer, preferably a polyethylene glycol / peptide polymer as defined above, and a lipid component, preferably a lipidoid component, more preferably a lipidoid component.

[0364] In a preferred embodiment of the second aspect, the at least one artificial RNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with a polymeric support, preferably with a polyethylene glycol / peptide polymer as defined above, and a lipidoid component, wherein the lipidoid component is a compound of formula A, wherein - R A independently selected at each occurrence from an unsubstituted, cyclic or acyclic, branched or unbranched aliphatic C 1-20 group; of a substituted or unsubstituted, cyclic or acyclic, branched or unbranched heteroaliphatic C 1-20 group; a substituted or unsubstituted aryl; a substituted or unsubstituted heteroaryl; - R5 is independently selected at each occurrence from an unsubstituted, cyclic or acyclic, branched or unbranched aliphatic C 8-16 ; a substituted or unsubstituted aryl; or a substituted or unsubstituted heteroaryl; - each occurrence of x is an integer from 1 to 10; - each occurrence of y is an integer from 1 to 10; or a pharmaceutically acceptable salt thereof.

[0365] In a preferred embodiment, the lipidoid component is 3-C12-OH according to formula B.

[0366] In preferred embodiments, the peptide polymer comprising lipidoid 3-C12-OH as specified above is used to complex the RNA of the first aspect and optionally the further artificial RNA of the second aspect to form complexes with an N / P ratio of about 0.1 to about 20, or of about 0.2 to about 15, or of about 2 to about 15, or of about 2 to about 12, wherein the N / P ratio is defined as the molar ratio of the nitrogen atoms of the basic groups of the cationic peptide or polymer to the phosphate groups of the artificial nucleic acid. Encapsulation / complexation in LNPs:

[0367] In preferred embodiments of the second aspect, the artificial mRNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed or associated with one or more lipids (e.g. cationic lipids and / or neutral lipids), thereby forming liposomes, lipid nanoparticles (LNPs), lipoplexes and / or nanoliposomes.

[0368] For compositions containing more than one artificial RNA construct defined herein (e.g., F-del and M2-1), these constructs can be formulated together, e.g., in LNPs, to form the respective composition.

[0369] Alternatively, more than one artificial RNA construct can be formulated separately and then combined to form the respective composition.

[0370] In this context, the terms “complexed” or “associated” refer to the essentially stable combination of the artificial RNA of the first aspect and, if applicable, the further artificial RNA of the second aspect with one or more lipids to form larger complexes or assemblies without covalent bonding.

[0371] The term "lipid nanoparticle," also known as "LNP," is not limited to a specific morphology and encompasses any morphology that arises when a cationic lipid and, optionally, one or more other lipids are combined, for example, in an aqueous environment and / or in the presence of RNA. Thus, a liposome, a lipid complex, a lipoplex, and the like fall within the scope of a lipid nanoparticle (LNP).

[0372] Accordingly, in preferred embodiments of the second aspect, the artificial mRNA of the first aspect and optionally the further artificial RNA of the second aspect is complexed with one or more lipids, thereby forming lipid nanoparticles (LNPs).

[0373] LNPs typically consist of a cationic lipid and one or more excipients selected from neutral lipids, charged lipids, steroids, and polymer-conjugated lipids (e.g., PEGylated lipid). The RNA may be encap...

Claims

[1] A pharmaceutical composition comprising at least one artificial mRNA, wherein the at least one artificial mRNA comprises: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence functionally linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from an RSV fusion protein (F), wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97% identical to SEQ ID NO: 4957, wherein the at least one antigenic peptide or protein is an F protein with a deleted C-terminus (F-del), and wherein the RSV F protein comprises a DSCav1 mutation (S155C, S290C, S190F and V207L), wherein the RSV F protein comprises the two subunits F2 and F1 in a single polypeptide chain, where F2 and F1 are connected via a linker element, where the F2 linker F1 protein aa104-aa144 is missing, wherein the RSV F protein includes at least one further mutation selected from A149C and Y458C, where the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO:

68. [2] The pharmaceutical composition according to claim 1, wherein the RSV F protein is stabilized in the prefusion conformation. [3] The pharmaceutical composition according to claim 1 or 2, wherein the linker element is a GS linker. [4] The pharmaceutical composition according to any of the preceding claims, wherein the F-protein with deleted C-terminus (F-del) is missing amino acids 554-574, wherein the position of an amino acid residue and the numbering refer to the position of the corresponding amino acid residue in SEQ ID NO:

68. [5] The pharmaceutical composition according to any one of the preceding claims, wherein the F protein with deleted C-terminus (F-del) lacks the C-terminal cytoplasmic portion. [6] The pharmaceutical composition according to any one of the preceding claims, wherein the F protein contains a transmembrane domain. [7] The pharmaceutical composition according to any one of the preceding claims, wherein the RSV F protein comprises the mutations A149C and Y458C. [8] The pharmaceutical composition according to any one of the preceding claims, wherein the RSV-F protein is F-del_DSCav1_mut4. [9] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one coding sequence encodes at least one amino acid sequence which is at least 98% or 99% identical to SEQ ID NOs: 2005, 2743, 3481, 4957 or 16523. [10] The pharmaceutical composition according to any of the preceding claims, wherein the at least one coding sequence encodes at least one amino acid sequence which is at least 99% identical to SEQ ID NOs: 4957. [11] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one coding sequence comprises at least one of the nucleic acid sequences that are identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90% identical to SEQ ID NOs 2006-2013, 2744-2751, 3482-3489, 4958-4965, 16524-16531, 21370, 21372, 21374, 21376, or 21402. [12] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one coding sequence comprises at least one nucleic acid sequence that is identical or at least 90% identical to SEQ ID NO: 4958. [13] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one coding sequence comprises at least one nucleic acid sequence that is identical or at least 90%, 91%, 92%, or 93% identical to SEQ ID NO: 4962. [14] The pharmaceutical composition according to any of the preceding claims, wherein the at least one coding sequence is a codon-modified coding sequence, wherein the codon-modified coding sequence is selected from C-maximized coding sequence, CAl-maximized coding sequence, coding sequence adapted to human codon use, G / C content modified coding sequence and G / C-optimized coding sequence or any combination thereof. [15] The pharmaceutical composition according to claim 14, wherein the Codon Adaptation Index (CAI) of the at least one coding sequence is at least 0.

9. [16] The pharmaceutical composition according to claim 14 or 15, wherein the Codon Adaptation Index (CAI) of the at least one coding sequence is at least 0.

95. [17] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA comprises a 5'-cap structure. [18] The pharmaceutical composition according to claim 17, wherein the 5'-cap structure is a cap1 structure. [19] The pharmaceutical composition according to any of the preceding claims, wherein the at least one artificial mRNA comprises at least one poly(A) sequence, preferably comprising 30 to 150 adenosine nucleotides, more preferably 100 adenosine nucleotides and / or at least one poly(C) sequence, preferably comprising 10 to 40 cytosines. [20] The pharmaceutical composition according to claim 19, wherein the at least one poly(A) sequence comprises 100 adenosine nucleotides. [21] The pharmaceutical composition according to one of the preceding claims, wherein all uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ). [22] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA comprises the following elements (a) a Cap1 structure, (b) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR) and (c) at least one coding sequence comprising at least one nucleic acid sequence that is at least 90% identical to SEQ ID NOs: 4958 or 4962; (d) a poly(A) sequence comprising about 100 adenosine nucleotides, wherein all uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ). [23] The pharmaceutical composition according to any one of the preceding claims, wherein the RSV-F protein is expressed in cells and presented on the cell surface after administration of the pharmaceutical composition. [24] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA comprises an RNA sequence which is at least 80% identical to a nucleic acid sequence selected from the group consisting of 4966-4967, 4974-4975, 4990, 4998, 4999, 5014, 5015, 5022, 5023, 5030, 5038, 5046, 5047, 5054, 5062, 5063, 5070, 5071, 5078, 5079, 5086, 5087, 5094, 5110, 5111, 5126, 5134, 5135, 5142, 5143, 5150, 5151, 5158, 5159, 5166, 5167, 5182, 5183, 5190, 5191, 5206, 5207, 5214, 5215, 5222, 5254, 5270, 5302, 5310, 5318, 5319, 21454-21456 and 21600-21602. [25] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA comprises an RNA sequence that is at least 80% identical to SEQ ID NO: 5318. [26] The pharmaceutical composition according to one of the preceding claims, wherein the at least one artificial mRNA is complexed with one or more lipids, thereby forming lipid nanoparticles (LNPs). [27] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA of LNPs is comprised. [28] The pharmaceutical composition according to any one of the preceding claims, wherein the at least one artificial mRNA or part thereof is associated or complexed with LNPs. [29] The pharmaceutical composition according to one of claims 26-28, wherein the LNP comprises one or more neutral lipids and / or a steroid. [30] The pharmaceutical composition according to claim 29, wherein the neutral lipid is selected from the group comprising distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE) and dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), 16-O-monomethyl-PE, 16-O-dimethyl-PE, 18-1-trans-PE, 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE) and 1,2-dielaidoyl-sn-glycero-3-phosphoethanolamine (transDOPE). [31] The pharmaceutical composition according to claim 29 or 30, wherein the neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and wherein the molar ratio of the cationic lipid to DSPC is optionally in the range of about 2:1 to 8:

1. [32] The pharmaceutical composition according to claim 29, wherein the steroid is cholesterol, and wherein the molar ratio of the cationic lipid to cholesterol is optionally in the range of about 2:1 to 1:

1. [33] The pharmaceutical composition according to any one of claims 26 to 32, wherein the LNP essentially consists of (i) at least one cationic lipid; (ii) a neutral lipid, preferably as defined in any one of claims 29 to 31; (iii) a steroid, preferably as defined in claim 32; and (iv) a PEG lipid, e.g. PEG-DMG, wherein (i) to (iv) are present in a molar ratio of approximately 20-60% cationic lipid, 5-25% neutral lipid, 25-55% sterol and 0.5-15% PEG lipid. [34] A pharmaceutical composition comprising at least one artificial mRNA, wherein the at least one artificial mRNA (a) a Cap1 structure; (b) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); (c) at least one coding sequence encoding at least one antigenic peptide or protein derived from an RSV fusion (F) protein, wherein the at least one coding sequence encodes at least one amino acid sequence that is at least 97%, 98% or 99% identical to SEQ ID NO: 4957, wherein the at least one coding sequence includes at least one nucleic acid sequence that is at least 90%, 91%, 92%, 93% identical to SEQ ID NO: 4958 or 4962; (d) a poly(A) sequence comprising about 100 adenosine nucleotides; wherein all uracil nucleotides are replaced by N1-methylpseudouridine (m1ψ) nucleotides; wherein the at least one artificial mRNA is contained in LNPs. [35] The pharmaceutical composition according to claim 34, wherein the RSV fusion (F) protein comprises the following mutations: S155C, S290C, S190F, V207L, A149C, and Y458C, and wherein the position of an amino acid residue and the numbering correspond to the position of the corresponding amino acid in SEQ ID NO:

68. [36] A vaccine comprising the pharmaceutical composition according to any one of claims 1 to 33 or according to claim 34 or 35. [37] The vaccine according to claim 36, wherein the vaccine elicits an adaptive immune response and / or a T-cell immune response against RSV. [38] A kit or kit of parts comprising the pharmaceutical composition according to any one of claims 1 to 33 or according to claim 34 or 35, and / or the vaccine according to claim 36 or 37, optionally comprising a liquid solubilizing agent, and optionally technical instructions with information on the administration and dosage of the components. [39] The pharmaceutical composition according to any one of claims 1 to 35, the vaccine according to claim 36 or 37, or the components of the kit or kit of parts according to claim 38 for use as a drug. [40] The pharmaceutical composition according to any one of claims 1 to 35, the vaccine according to claim 36 or 37, or the components of the kit or kit of parts according to claim 38 for use in the treatment or prophylaxis of an infection with RSV in a subject or of a disorder associated with such an infection. [41] The pharmaceutical composition according to any one of claims 1 to 35, the vaccine according to claim 36 or 37, or the components of the kit or kit of parts according to claim 38 for use in a method for reducing the RSV titer in nasal tissue of a subject who has an RSV infection. [42] The pharmaceutical composition according to any one of claims 1 to 35, the vaccine according to 36 or 37, or the components of the kit or kit of parts according to claim 38 for use in a method for reducing the RSV titer in lung tissue of a subject who has an RSV infection. [43] The pharmaceutical composition according to any one of claims 1 to 35, the vaccine according to claim 36 or 37, or the components of the kit or kit of parts according to claim 38 for use according to any one of claims 39-42, wherein the subject is a human subject, preferably a newborn, a pregnant woman, a breastfeeding woman, an elderly and / or an immunocompromised human subject.

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